Click here to load reader

Statistical methods for single-cell RNA sequencing data...Statistical methods for single-cell RNA sequencing data ... XX XXXXXXX X XXX X XXX XXX X XXX XXX XX X X XX XXX X XX XX XX

  • View
    1

  • Download
    0

Embed Size (px)

Text of Statistical methods for single-cell RNA sequencing data...Statistical methods for single-cell RNA...

  • CK BioC 2017

    Statistical methods for single-cell RNA sequencing data

    Department of Biostatistics and Medical InformaticsUniversity of Wisconsin-Madison

    http://www.biostat.wisc.edu/~kendzior/

  • CK BioC 2017

    Single-cell vs. bulk RNA-seq

    Heterogeneous Homogeneous Sub-population

  • CK BioC 2017

    Features of single-cell RNA-seq data

    § Abundance of zeros, increased variability, complex distributions

    Bacher and Kendziorski, Genome Biology, 2016.Log (variance) Number of clusters

    Prop

    ortio

    n of

    zer

    osD

    ensi

    ty

    Expression Group

    Cou

    nt

  • CK BioC 2017

    § Normalization

    § Technical vs. biological zeros

    § Clustering; Identifying sub-populations

    § De-noising

    § Adjusting for technical variability

    § Adjusting for biological variability (oscillatory genes)

    § Identifying and characterizing differences in gene-specific expression distributions (aka. identifying differential distributions)

    § Pseudotime reordering

    § Network reconstruction

    Challenges in scRNA-seq

  • CK BioC 2017

    § Normalization

    § Technical vs. biological zeros

    § Clustering; Identifying sub-populations

    § De-noising

    § Adjusting for technical variability

    § Adjusting for biological variability (oscillatory genes)

    § Identifying and characterizing differences in gene-specific expression distributions (aka. identifying differential distributions)

    § Pseudotime reordering

    § Network reconstruction

    Challenges in scRNA-seq

  • CK BioC 2017

    Variability induced by oscillatory genes is substantialin single-cell RNA-seq and can mask effects of interest

    We developed an approach called Oscope to identify and characterize oscillations in single-cell RNA-seq experiments

  • CK BioC 2017

    Oscope identifies oscillatory genes in unsynchronized single-cell RNA-seq experiments

    Leng et al., Nature Methods, 2015

  • CK BioC 2017

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    −0.50

    0.5

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    −0.50

    0.5

    ●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    −0.50

    0.5

    cell80

    cell25

    cell1

    cell3

    cell2

    cell4

    Oscope'(Oscilloscope):'A'sta0s0cal'pipeline'for'iden0fying'oscillatory'gene'sets'

    Module'1:'Screening'for'candidate'oscillators'(Paired

  • CK BioC 2017

    Time

    ●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●

    −0.50

    0.5

    ●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●X

    X

    ●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●−0.5

    00.5

    ●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●X

    X

    ●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●

    −0.50

    0.5

    ●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●XX

    ●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●−0.5

    00.5

    ●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●X

    X

    t0,1

    t0,2

    t0,3

    t0,4 T

    Oscope: Identifying oscillatory genes using scRNA-seq

    t4

    t1

    cell 1

    cell 2

    cell 3

    cell 4

    gene 1gene 2−0.5

    00.5

    X

    X

    −0.50

    0.5 X

    X

    −0.50

    0.5 XX

    −0.50

    0.5 X

    X

    t0,1

    t0,2

    t0,3

    t0,4 T

    Collection time

    t2

    t3

    Oscillation time

    t4

    X

    X

    X

    X

    t1

    XX

    t3

    X

    X

    t2

    XX

    XX X

    XXX

    −0.50

    0.5

    cell1

    (t1

    )

    cell2

    (t2

    )ce

    ll3 (

    t3)

    cell4

    (t4

    )

    XXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXXX

    XXXXXXX

    XXXXXXXX

    XXXXXXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXX

    XX

    X X

    XXX

    −0.50

    0.5

    cell1

    (t1)

    cell2

    (t2)

    cell3

    (t3)

    cell4

    (t4)

    XXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXXX

    XXXXXXX

    XXXXXXXX

    XXXXXXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXXXXXX

    XXXXXXXXX

    XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

    XXXXXXXX

    XXX

    −1.0

    −0.5

    0.0

    0.5

    1.0Scaled_Expression

    cell1

    cell3

    cell4

    cell2

    ...

    ......

    gene1

    gene2

    Cyclic order

    Base cycle Base cycle

  • CK BioC 2017

    § Given a list of cities, only know distances between each pair of cities

    Distance (mile)

    Madison-Chicago 148

    Madison-Iowa City 175

    Madison-Atlanta 847

    Madison-Minneapolis 273

    Chicago-Iowa City 223

    Chicago-Atlanta 716

    … …

    § In our case, for each gene cluster, we want to find an optimal “route” through all cells and return to the first cell

    § The optimal route will minimize expression differences between observed and baseline oscillation−1.0

    −0.5

    0.0

    0.5

    1.0Scaled_Expression

    cell1

    cell3

    cell4

    cell2

    ...

    ...

    ...

    gene1

    gene2

    § Goal is to find an optimal route that visits each city exactly once and returns to the origin city

    § The optimal route will minimize overall distance travelled

    Oscope: Connection to TSP

  • CK BioC 2017

    § Whitfield data: microarray time course of HeLa cells synchronized for cell cycle. 48 samples; one every hour (~3 cell cycles).

    § Applied Oscope on Whitfield data with permuted sample order

    − Top cluster has 69 genes (65 of 69 validated as oscillating in Whitfield).

    Oscope: Results from Whitfield data

    Scal

    ed E

    xpre

    ssio

    n

    ENI Reconstructed Order0 10 20 30 40 50

    ●●●●●●

    ●●●

    ●●●●●●●●

    ●●●●●

    ●●

    ●●●●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●●●

    CCNE1

    ●●●

    ●●●●●

    ●●

    ●●●●●●●●

    ●●●●

    ●●●●●●●●●●

    ●●●●●●●●●●

    ●●

    ●●●●

    PLK

    ●●●●●●●

    ●●●

    ●●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●

    H2AFX

    ●●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●●●

    ●●●●

    ●●

    ●●●●●

    ●●●●●●●●

    ●CDC6

    −0.5

    0.51

    −1

    0

    1

    −0.6

    0

    0.6

    −0.5

    0.50

    0

    −0.5

    0.51

    0 10 20 30 40 50

    ●●●●●

    ●●●●

    ●●●●

    ●●●

    ●●

    ●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●

    ●●●

    ●●●●●●●●

    CCNE1

    −1

    0

    1

    ●●●●●

    ●●●

    ●●●●

    ●●

    ●●●

    ●●●●●●

    ●●●●●●●●

    ●●●●●●●●●●

    ●●●

    ●●●●

    PLK−0.6

    0

    0.6

    ●●●●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●●●

    ●●●

    ●●

    ●●●●●

    ●●●●●●●

    ●●●●●●●

    ●●●

    H2AFX

    −0.5

    0.5 ●●●●

    ●●●

    ●●●●●●

    ●●●●●

    ●●

    ●●●●●●

    ●●

    ●●●●●

    ●●●●●●●●

    ●●●●

    ●●●

    CDC6

    0

    0

    Scal

    ed E

    xpre

    ssio

    n

    Known Time Course Order

  • CK BioC 2017

    Recovered order

    0

    1,000

    2,000

    0 50 100 150 200

    ●●

    ●●●●●●●

    ●●●●

    ●●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●●●●

    ●●●●●●

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●●●●●●

    ●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●

    ●●

    ●●

    ●●●●

    TPX20

    2,0004,000

    ●●●

    ●●●●

    ●●●

    ●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●

    ●●●●●

    ●●

    ●●●●

    ●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●●●●●●●

    ●●●●●

    ●●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●●●●

    ●●●

    ●●●

    ●●●●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●

    ●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●●

    CCNB10

    15,000

    ●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●

    ●●●●●●●●●●●●●●●

    ●●●●●●

    ●●●●●●

    ●●●●●●●

    ●●

    ●●●●●●

    ●●●

    ●●

    ●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    KPNA20

    1,500

    3,000

    ●●●●●●●●●●●●●●●●

    ●●●

    ●●

    ●●●●●

    ●●●

    ●●●●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●●

    ●●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●●●●●●

    NUSAP1

    30,000

    6,000

    Scal

    ed E

    xpre

    ssio

    n

    1

    0.5

    -0.5

    -1

    1

    0

    -1

    1

    0

    -1

    0.8

    0

    -0.8

    ENI Reconstucted Order

    1

    0.5

    -0.5

    -1

    1

    0

    -1

    1

    0

    -1

    0.8

    0

    -0.8

    Scal

    ed E

    xpre

    ssio

    n

    ENI Reconstructed Order

    Oscope: Results from H1 hESCs (with Thomson lab)

    § Oscope applied to 213 H1 hESCs identified a 29 gene group − 21 of 29 genes annotated as cell-cycle by GO.

    § To investigate this group, Oscope was reapplied to 460 H1 hESCs− 213 unlabeled and 247 FUCCI labeled (cell cycle phase is known).

    G1G2/M

    S

    Cells are characterized as G1, S, G2/M

    CC phase assignment is masked and Oscope is applied to the 29 genes

    An accurate reconstructed order will clearly separate the three CC phases

  • CK BioC 2017

    Oscope identifies potential artifact in Fluidigm C1 platform

    ID

    Expression

    01000

    0 50 100 150 200

    ●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●●

    ●●●

    ●●

    ●●●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●

    ●●

    GARS

    01500

    ●●

    ●●

    ●●●●

    ●●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●●●●●●●●●●●●

    ●●

    ●●●●●●●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●

    ●●●●

    ●●

    ●●●●

    ●●●●●

    ●●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●●●●

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●●●●

    STRN3

    0200500

    0 50 100 150 200

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●

    ●●

    ●●

    ●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●●●●

    ●●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●●

    ●●●

    ●●●●●

    ●●

    ●●●●●

    ●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●

    WDR18

    0400800

    ●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●

    RALY0

    400800

    0 50 100 150 200

    ●●●

    ●●

    ●●●●●●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●●●

    ●●

    ●●●●●●●

    ●●●

    ●●

    ●●●●●●●●●●

    ●●

    ●●●●●●

    ●●●●

    ●●●●

    ●●●●●●●

    ●●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●●●●●●●

    ●●

    ●●●●●●●●●●●●

    ●●●

    ●●●●●

    ●●

    VTI1B

    0500

    1500

    ●●●

    ●●

    ●●

    ●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●●●●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●

    ●●●

    PPP2CA

    0500

    1500

    ●●●●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●●

    ●●●●

    ●●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●●●●

    ●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●●

    ●●●●●●●●●

    ●●●●●●●●●●●

    ●●

    ●●●●●●

    ●●●●●

    ●●●●●●●●●

    PIGY

    0200

    500

    ●●

    ●●●●●●●●●

    ●●●

    ●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●

    ●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●

    ●●●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●●●

    ●●●●●●●●●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●●

    ●●

    TBCB

    0200500

    ●●●●

    ●●●●●●●●●●

    ●●●

    ●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●

    ●●

    ●●●●●●●

    ●●●

    ●●●●●

    ●●●

    ●●●●●●●●●●

    ●●●

    ●●

    ●●●●

    ●●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●

    ●●●●●●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    APOE

    050150

    ●●●

    ●●●

    ●●

    ●●●●●

    ●●●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●●●●●

    ●●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●●●●●●●●●●●

    ●●●●●

    ●●●

    ●●●●●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●

    ●●●

    ●●●●●

    ●●●●●●

    ●●

    ●●●●●●

    ●●●●

    ●●

    ●●●●●●●●●

    DUSP23

    04000

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●●●●●●●●

    ●●●●●●

    ●●●

    ●●

    ●●●●●●●●

    ●●●

    ●●●●●●

    ●●

    ●●●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●●

    ●●

    ●●

    ●●●●●●

    ●●●●●

    ●●●●●●●●●

    ●●●●●●●●●

    ●●●

    ●●●●●●

    ●●●

    SOX11

    010003000

    ●●●

    ●●●

    ●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●

    ●●●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●

    CNN3

    0100250

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●●

    ●●

    ●●●

    ●●●●●●●●●●●●

    ●●●●●●

    ●●

    ●●●●●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●

    ●●●

    ●●●●●

    ●●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●●●●

    ●●●●

    ●●

    ●●●●●●

    ●●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●●●

    ●●●

    C16orf13

    0400800

    ●●●●

    ●●

    ●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●

    ●●●

    ●●

    ●●●

    ●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●●●

    ●●

    ●●●●

    ●●●●●●

    ●●

    ●●●●●●●●●●●●●●

    ●●●●●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●●

    ●●●●●●●●

    ●●●●●

    ●●●●

    ●●

    ●●●●●●

    ●●●●●

    ●●●

    ●●●●●

    ●●

    ●●

    ●●

    ADI1

    05001500

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●

    ●●●●●

    ●●●●●

    ●●

    ●●●●

    ●●

    ●●●●●

    ●●●●●●

    ●●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●

    ●●●

    ●●●

    ●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●

    ●●●●

    ●●●●●

    ●●●●

    KRT19

    0200400

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●●●●●●

    ●●●●●

    ●●●●●

    ●●●●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●●

    ●●●

    ●●●●

    ●●●●●

    ●●●

    ●●●●●●

    ●●●

    ●●●●●●●●

    MRPS26

    0500

    1500

    ●●●●●●

    ●●

    ●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●

    ●●●●●●●

    ●●●●

    ●●

    ●●

    ●●●●

    ●●●●

    ●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●

    ●●●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●

    PDCD6

    0400800

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●●●

    ●●

    ●●●●●●●●●●●●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●●●●●

    ●●

    ●●●●

    ●●●●

    ●●●●●●

    ●●●●●●

    ●●

    ●●●●●●●●●●●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●

    ●●●

    ●●

    ●●●●

    ●●●●●●●●

    ●●●

    ●●●

    ●●

    ●●●●●●●●

    ●●●

    ●●

    ●●●

    ●●

    SUMO3

    0100

    300

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●●●

    SNF8

    02000

    ●●

    ●●

    ●●●

    ●●●●●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●

    ●●●●●●

    ●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●●●

    ●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●●●●

    ●●●●●●●●●●

    ●●●●●●●●●●

    ●●●

    ●●●●●●●●

    ●●●●●●

    ●●

    ●●●

    TUBB2A

    0200

    600

    ●●●

    ●●●●●●●●●

    ●●●●●●

    ●●

    ●●●●

    ●●

    ●●●●

    ●●●●●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●●●●●●●●●

    ●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●

    ●●●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●●●

    ●●●●●●●

    ●●●●

    CLTA

    050150

    ●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●

    ●●●

    ●●●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●

    ●●

    ●●●●●●●●●●●

    ●●●●●●●●●●●●●●

    CYBA

    02060

    100

    ●●●

    ●●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●●●●●●

    ●●

    ●●●●●●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●●

    ●●

    ●●●●●

    ●●●●

    ●●

    ●●●

    TRAPPC5

    050

    150

    ●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●●●●●●●

    ●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●

    ●●●

    ●●●●●●●●

    ●●

    ●●●●●●

    ●●●●●●●●●●●●

    ●●●●●●●●

    ALKBH7

    0200

    600

    ●●●

    ●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●

    ●●●●

    ●●●●●

    ●●

    MRPL12

    0200

    500

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●●●

    ●●●●

    ●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●

    ●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●

    ●●●●●●●●●●

    ●●●

    ●●●●●

    ●●

    TMEM93

    0500 ●●●●

    ●●●

    ●●

    ●●

    ●●●●●●

    ●●●●●●●

    ●●

    ●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●●

    ●●●●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●●

    ●●

    EIF4EBP1

    0500

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●●●●●●

    ●●●

    ●●●

    ●●●●●●●

    ●●●

    ●●●●●●●●●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●

    ●●●●●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●

    ●●●●●

    ●●●●

    ●●

    ●●●

    ●●●●●●●

    ●●●●

    ●●●●●●●

    LSM4

    0500

    1500

    ●●●●

    ●●

    ●●●●●●●●

    ●●●●●

    ●●●●●●●

    ●●●●

    ●●●●●●●●●●●●●●

    ●●●

    ●●●●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●●

    ●●

    ●●●●

    ●●●●●●

    ●●●●●●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●●●●●

    ●●●●●

    ●●

    ●●●●●●●

    ●●

    ●●●

    ●●●●

    ●●●●

    ●●

    ●●●

    ●●

    HNRNPM

    0200400

    ●●●●

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●●

    ●●●●

    ●●●

    ●●●●●

    ●●●

    ●●●●●●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●●●

    ZSCAN10

    0400800

    ●●

    ●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●

    ●●●●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●

    ●●●●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●

    ●●●

    ●●

    ●●●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    DUT

    0200500

    ●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●●

    ●●●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●●●●

    ●●●●

    ●●

    ●●

    ●●●●●●●●●

    ●●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●●●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●●●

    ●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●

    ISCU

    5002000

    ●●

    ●●

    ●●●●●●

    ●●●

    ●●●●●

    ●●●

    ●●●●

    ●●●●●●●●●●

    ●●●●●●●

    ●●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●●

    ●●●●

    ●●

    ●●●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●●●●

    ●●●●●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●●●●●

    ●●●

    ●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●

    ●●

    ●●●●●●●

    ●●

    RPS2

    0200500

    ●●

    ●●●

    ●●●

    ●●●●

    ●●

    ●●●●●●●

    ●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●●

    ●●

    ●●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●●

    ●●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●●●●●

    ●●●●●●

    ●●●●

    ●●●●●●●●●●●

    ●●●

    ●●

    NOL7

    0100

    250

    ●●●●●

    ●●●

    ●●●●●●●●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●●●

    ●●●●●●

    ●●●●●●

    ●●●●●●●●●

    ●●●●●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●

    SCAND1

    0500

    1500

    ●●

    ●●●●

    ●●●●

    ●●●●●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●●●●●●●

    ●●●●●

    ●●●

    ●●●

    ●●

    ●●●

    ●●●

    ●●●

    ●●●●●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●●●

    ●●●●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●●●

    ●●●

    ●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●●

    ●●●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●

    EIF5A0200

    600

    ●●●

    ●●

    ●●●●

    ●●

    ●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●●

    ●●●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●●●●

    ●●

    ●●●●

    ●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●●●

    ●●●

    ●●

    ●●●●

    ●●●●●●●

    ●●●●●●

    ●●●●

    ●●●

    ●●●

    ●●●●

    RANBP1

    0200400

    ●●●

    ●●

    ●●●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●●

    ●●

    ●●●●●●●

    ●●●●●●●●

    ●●

    ●●

    ●●

    ●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●●●●●●●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●

    ●●●●●

    TNFRSF12A

    0400800

    ●●

    ●●

    ●●●

    ●●●●●●

    ●●

    ●●

    ●●●●●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●●●●●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●

    ●●●●

    ●●●●●●●●

    ●●●●●●

    ●●

    ●●●●●●●

    ●●●●●

    ●●●●●●●●

    ●●

    ●●●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●●●●●●●●●●●

    ●●●

    ●●●●

    ●●●●

    ●●●●

    ●●

    ●●●

    ●●●

    ●●●●●●●

    GCSH

    0100

    300

    ●●

    ●●●

    ●●

    ●●

    ●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●●

    ●●

    ●●

    ●●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●

    ●●●●●

    ●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●

    ●●●●

    ●●

    ●●●

    ●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●●

    ●●●●●●

    ●●●●●

    ●●●●●●

    ●●●●●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●●

    ●●

    C19orf10

    010003000

    ●●

    ●●

    ●●●●●●●●●●●●

    ●●●●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●

    ●●●●●

    ●●●●

    ●●●●

    ●●

    ●●●●●

    ●●

    ●●●●●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●

    ●●●●

    ●●●●●●●●●

    ●●●●●

    ●●●●●●●●●●●●●

    ●●●●●●●●

    ●●●

    ●●●

    ●●●

    ●●●●

    ●●●●

    PDIA3

    0400800

    ●●

    ●●●●

    ●●●

    ●●●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●●●●●●●●●

    ●●●●●●●●●●●●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●●

    ●●●●●●

    ●●●●●●●●●●●●●●

    ●●●●

    ●●●●

    ●●

    ●●●

    ●●●●●

    ●●

    ●●●●●●

    ●●

    ●●●

    ●●●●●●

    ●●●●

    ●●

    ●●●

    ●●●●●●

    ●●●●●●●●●●●

    ●●

    ●●

    ●●

    LSM2

    050

    100 ●

    ●●

    ●●

    ●●

    ●●

    ●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●●●

    ●●

    ●●●●●●●

    ●●●

    ●●●

    ●●●●

    ●●●●●●●●●●●

    ●●●●●●●

    ●●●●

    ●●

    ●●●●●●●●●●●

    ●●●●●●●

    ●●●●

    MRPL41

    0400800

    ●●

    ●●●

    ●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●

    ●●●●●

    ●●●●●●●●●●●●●●●

    ●●●

    ●●●●●●●●●●●●●●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●●

    ●●●●●●

    ●●●

    ●●●

    ●●●●●

    ●●●●●

    ●●●●

    ●●●

    ●●

    ●●●●●

    ●●●●●●●●●

    ●●●●●●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●●

    COX5A

    010003000

    ●●●●

    ●●●

    ●●●●

    ●●

    ●●●●●●

    ●●●●

    ●●●●●

    ●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●●●●

    ●●●

    ●●●●●●●

    ●●●●●

    ●●●

    ●●

    ●●

    ●●●●

    ●●

    ●●

    ●●

    ●●●●

    ●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●

    ●●●

    ●●●●●●●●●

    ●●

    ●●

    ●●

    MIF

    02000

    ●●

    ●●

    ●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●●●●●●●●●●

    ●●

    ●●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    PFN1

    0 50 100 150 200

    0400800

    ●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●

    ●●●●●●●●

    ●●●●●●●●●

    ●●

    ●●●

    ●●●●●

    ●●

    ●●

    ●●●●●

    ●●

    ●●

    ●●●

    ●●●

    ●●

    ●●●

    ●●●●●●●●●

    ●●●

    ●●●

    ●●●

    ●●●●●●

    ●●●

    ●●

    ●●●

    ●●

    ●●

    ●●

    ●●

    ●●

    ●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●

    ●●●●●

    IMP3

    02000

    ●●

    ●●●●

    ●●

    ●●●●●●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●

    ●●●●●●●●

    ●●●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●

    ●●

    ●●

    ●●●●●

    ●●●

    ●●

    ●●●

    ●●●●●●●●●●●●●●●●

    ●●●

    ●●●

    ●●●

    ●●

    ●●

    ●●●

    ●●

    ●●●

    ●●●●

    ●●●●●●

    ●●●

    ●●●●●●●●●●●●

    ●●●●●●●●●

    ●●●●

    ●●

    ●●●●●●●●●

    RPL13

    0 50 100 150 200

    01000

    2500

    ●●

    ●●

    ●●●●

    ●●●●●●

    ●●●●●●

    ●●

    ●●●●●●●●●●●●●

    ●●●●●●●●●●●●

    ●●●

    ●●●

    ●●●●

    ●●●

    ●●●

    ●●

    ●●●●●●●●●●

    ●●●●●●

    ●●●●●●

    ●●●●●●●●●●●●●

    ●●

    ●●●

    ●●●●

    ●●●●●

    ●●●●

    ●●●●●●●●●●●●

    ●●●●●●●

    ●●●●●

    ●●●●●●●●●●●●●●

    ●●

    ●●

    ●●●

    EIF3F

    02000

    5000

    ●●●

    ●●●●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●●●

    ●●

    ●●

    ●●

    ●●●

    ●●●

    ●●●●●●●●●●●●●●●●●

    ●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●

    ●●

    ●●

    ●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●●

    ●●●●●●●

    ●●●●●●●●●●●●●●●●●●●●●●

    FTH1

  • CK BioC 2017

    Schematic of Fluidigm’s C1 platform

    !

    C48C47C46C45C44C43C42C41C40C39C38C37C36C35C34C33C32C31C30C29C28C27C26C25C24C23C22C21C20C19C18C17C16C15C14C13C12C11C10C09C08C07C06C05C04C03C02C01

    C96C95C94C93C92C91C90C89C88C87C86C85C84C83C82C81C80C79C78C77C76C75C74C73C72C71C70C69C68C67C66C65C64C63C62C61C60C59C58C57C56C55C54C53C52C51C50C49

    (P93)(P92)(P91)(P87)(P86)(P85)(P81)(P80)(P79)(P75)(P74)(P73)(P69)(P68)(P67)(P63)(P62)(P61)(P57)(P56)(P55)(P51)(P50)(P49)(P43)(P44)(P45)(P37)(P38)(P39)(P31)(P32)(P33)(P25)(P26)(P27)(P19)(P20)(P21)(P13)(P14)(P15)(P07)(P08)(P09)(P01)(P02)(P03)

    (P94)(P95)(P96)(P88)(P89)(P90)(P82)(P83)(P84)(P76)(P77)(P78)(P70)(P71)(P72)(P64)(P65)(P66)(P58)(P59)(P60)(P52)(P53)(P54)(P48)(P47)(P46)(P42)(P41)(P40)(P36)(P35)(P34)(P30)(P29)(P28)(P24)(P23)(P22)(P18)(P17)(P16)(P12)(P11)(P10)(P06)(P05)(P04)

    C46

    C43

    C40

    C37

    C34

    C31

    C28

    C25

    C24

    C21

    C18

    C15

    C12

    C09

    C06

    C03

    C47

    C44

    C41

    C38

    C35

    C32

    C29

    C26

    C23

    C20

    C17

    C14

    C11

    C08

    C05

    C02

    C48

    C45

    C42

    C39

    C36

    C33

    C30

    C27

    C22

    C19

    C16

    C13

    C10

    C07

    C04

    C01

    C96

    C93

    C90

    C87

    C84

    C81

    C78

    C75

    C70

    C67

    C64

    C61

    C58

    C55

    C52

    C49

    C95

    C92

    C89

    C86

    C83

    C80

    C77

    C74

    C71

    C68

    C65

    C62

    C59

    C56

    C53

    C50

    C94

    C91

    C88

    C85

    C82

    C79

    C76

    C73

    C72

    C69

    C66

    C63

    C60

    C57

    C54

    C51

    (P91)

    (P85)

    (P79)

    (P73)

    (P67)

    (P61)

    (P55)

    (P49)

    (P43)

    (P37)

    (P31)

    (P25)

    (P19)

    (P13)

    (P07)

    (P01)

    (P92)

    (P86)

    (P80)

    (P74)

    (P68)

    (P62)

    (P56)

    (P50)

    (P44)

    (P38)

    (P32)

    (P26)

    (P20)

    (P14)

    (P08)

    (P02)

    (P93)

    (P87)

    (P81)

    (P75)

    (P69)

    (P63)

    (P57)

    (P51)

    (P45)

    (P39)

    (P33)

    (P27)

    (P21)

    (P15)

    (P09)

    (P03)

    (P94)

    (P88)

    (P82)

    (P76)

    (P70)

    (P64)

    (P58)

    (P52)

    (P46)

    (P40)

    (P34)

    (P28)

    (P22)

    (P16)

    (P10)

    (P04)

    (P95)

    (P89)

    (P83)

    (P77)

    (P71)

    (P65)

    (P59)

    (P53)

    (P47)

    (P41)

    (P35)

    (P29)

    (P23)

    (P17)

    (P11)

    (P05)

    (P96)

    (P90)

    (P84)

    (P78)

    (P72)

    (P66)

    (P60)

    (P54)

    (P48)

    (P42)

    (P36)

    (P30)

    (P24)

    (P18)

    (P12)

    (P06)

    (e)

    (c)

    (d)

    (a)

    (b)

    ID

    qPC

    R F

    C0.

    01.

    02.

    0

    0 50 100 150 200

    !!!!!!!!!!!!!!!!!!!

    !!!!!

    !!

    !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

    !!

    !!!!!!

    !

    !

    !!!!!

    !!!

    !!!!!!!!!

    !!!!!!!!

    !

    !!!!

    !!

    !

    !!!!!!!!

    !!!!!!!!!!!!!!

    !

    !

    !

    !!

    !

    !

    !!!!!!

    !!

    !

    !

    !

    !

    !

    !

    !!

    !!

    !

    !!!!

    !!

    !

    !

    !

    !

    !!!

    !!

    !

    !!!!!!!

    !!

    !!!!!!!!!!!!!!!

    !!!!!!

    !!!!

    !!

    !

    !!!

    !

    !!

    !

    !

    !

    MIF0.0

    1.0

    2.0

    !

    !

    !!!!!!

    !!!!!!!!!!!!!!!!!!

    !!!!!!!!!!!!!!!!!!!!!!!!!!

    !!!!!!!!

    !!

    !!

    !!!

    !

    !!

    !

    !!

    !

    !!!

    !!!!!!!!

    !!!

    !!!!!!!

    !

    !!!!!

    !!!!!!!!

    !

    !

    !!!!!!!!

    !

    !!!

    !

    !

    !

    !!

    !!!!!!!

    !!!!

    !!

    !!

    !!!

    !!

    !!!!!!!!!!!!!!!!

    !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

    !!!!!!!!!!!!!

    PFN1

    ID

    FPKM

    0100

    300

    0 100 200 300

    !

    !!

    !!!

    !!

    !

    !

    !

    !!

    !

    !

    !!!!!

    !

    !!!!!!!!!!!!

    !

    !!!!!!!!!!!!!!!

    !

    !!!!

    !

    !!!!!

    !

    !

    !!

    !!!

    !!!!

    !

    !!

    !

    !

    !

    !!!

    !

    !

    !

    !!!!!!!!

    !

    !!!

    !!

    !!!

    !!!

    !

    !

    !

    !!!!!

    !

    !

    !

    !

    !!!!!!!

    !

    !

    !!!!

    !

    !

    !!!!!!!!!!!

    !

    !!!!!!!!

    !

    !!!!!!!!!

    !!

    !!!!

    !!

    !

    !

    !

    !

    !!!!!

    !

    !

    !

    !!

    !

    !!

    !

    !!!!

    !

    !

    !!

    !

    !

    !

    !

    !!!!!

    !!!

    !!!!!!

    !

    !

    !!

    !

    !!!

    !

    !

    !

    !!!!!!!!!!!!

    !

    !

    !

    !!!!!!!!

    !

    !!!!

    !

    !

    !

    !!!!!!!

    !

    !!!!!!!!!

    !

    !!!!

    !

    !!!!

    !

    !

    !!!!

    !

    !!

    !

    !

    !

    !

    !

    !

    !!!

    !

    !!

    !

    !

    !

    !!!!!!!!!!!

    !

    !

    !

    !

    !

    !

    !

    !!!!!!

    !

    !

    !!!!!!!!!!

    !

    !!

    !

    !!

    !

    !!!!!!!

    !!!!!!!!!!

    !!!!

    !

    !

    COX5A

    010002500

    !

    !!

    !

    !

    !!!

    !!

    !!

    !

    !

    !

    !

    !!!!!

    !

    !

    !!!!

    !

    !!!!

    !

    !

    !!

    !!!

    !!!!!!

    !

    !!!!!!!!!!!!!!!!!!!!!!!

    !!!!!!!!!

    !!

    !

    !

    !

    !!!!!

    !!

    !

    !!!

    !

    !

    !

    !

    !

    !!

    !

    !

    !!!

    !!

    !!

    !

    !

    !

    !

    !

    !!

    !

    !

    !

    !!

    !

    !

    !

    !

    !!

    !

    !

    !

    !

    !!!!!!!!!!!

    !!

    !!

    !

    !

    !

    !

    !

    !!

    !!!!

    !

    !!

    !

    !

    !

    !!!!!

    !!

    !!!

    !!

    !

    !!

    !!!

    !

    !

    !

    !

    !

    !

    !

    !!

    !

    !

    !

    !

    !!

    !!

    !!

    !

    !!

    !!

    !!

    !!

    !

    !

    !

    !

    !!!!!!!!!!!

    !

    !!!!!!!!

    !

    !!!!!!!!!!!!!!!!

    !

    !

    !

    !!

    !

    !

    !

    !!!

    !

    !!

    !!

    !

    !!

    !

    !

    !!!

    !

    !

    !!!

    !

    !

    !

    !

    !

    !!

    !

    !!

    !!

    !

    !

    !

    !!!!!!

    !

    !

    !!

    !!!!

    !

    !!!

    !

    !!!

    !

    !

    !

    !!!!!!!

    !!!!

    !!!!!

    !

    !!

    !

    !!!!!

    !!!

    !

    !!

    !!

    !!!!!!

    !

    !!

    !

    !!

    !!!

    !!

    RPL13

    0100250

    !

    !

    !

    !!

    !

    !

    !!!

    !

    !

    !

    !

    !

    !

    !!!

    !

    !

    !

    !!

    !

    !

    !

    !

    !!

    !

    !

    !

    !!!

    !

    !

    !

    !!!

    !!

    !

    !!!!!!!

    !!!!!!!!

    !

    !!!

    !!!!

    !

    !!!!!!!!!

    !!

    !!

    !!!

    !!!!

    !

    !

    !!

    !!

    !

    !!

    !

    !

    !!

    !

    !

    !

    !

    !

    !

    !

    !

    !!

    !

    !

    !

    !

    !

    !

    !

    !!

    !

    !

    !

    !

    !!!!!!

    !!!

    !

    !

    !

    !

    !!!

    !

    !!

    !!

    !

    !!

    !

    !

    !!

    !!

    !!!

    !

    !!

    !

    !

    !!!

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !!

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !

    !!

    !

    !!!

    !

    !

    !

    !!

    !

    !

    !!

    !

    !

    !

    !!!

    !

    !!

    !

    !

    !!

    !!

    !

    !!!!!

    !!

    !

    !

    !

    !!!!

    !

    !

    !!

    !!!

    !!!

    !

    !