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Supplementary Figure 1

Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

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Page 1: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 1

Page 2: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 2

Page 3: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 3

A

B

Page 4: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

A B

C

Supplementary Figure 4

Page 5: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 5

A

B

Page 6: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

AREG

HBEGF

EGF

ErbB3

EREG

TGFα

BTC

ErbB2

Mouse secondary

Goat secondary

Supplementary Figure 6

Page 7: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

7

Days

Co

nce

ntr

atio

n u

g/m

L

0.1

1

10

100

0 5 10 15 20 25 30

B07

0.1 mg/kg1 mg/kg10 mg/kg

Supplementary Figure 7

Page 8: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

10mg/kg Nonspecific IgG 10mg/kg MEDI3622

tEGFR

tErB3

ErB2(Tyr1112)

ErB2(Thr686)

EGFR(Tyr1086)

EGFR(Tyr845)

EGFR(Ser1070)

EGFR(Tyr1173)

Supplementary Figure 8

Page 9: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Nonspecific IgG #1

MEDI3622 #2MEDI3622 #1

Nonspecific IgG #2

Yes

Src

Supplementary Figure 9

Page 10: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 10

Page 11: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 11

Page 12: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 12

A

B

Page 13: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 12

C

Page 14: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 13A B

C D

Page 15: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Figure 14

Page 16: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Table 1Cell line

% Max Inhibition MEDI3622

% Max Inhibition cetuximab

786-0 0.2 0.3

A549 3.4 6.1

BxPC3 16 3

C2BBE1 19.7 12

CakiI 19.2 25.6

Cal27 20.7 21.7

CCK81 8.7 77.5

Detroit 32.6 32.9

Du145 13.8 9.9

Fadu 16.5 35.2

H292 33.4 53.8

H358 41.5 65.9

HSC3 0 4.8

HT29 0 5.1

KYSE30 5.2 7.7

KYSE450 7.6 43.3

NUGC4 65.8 91.5

OE21 25 33.9

OE33 0.5 0.6

panc0203 16.4 5

PLCPRF5 30.9 48.2

SCC4 32.1 40.7

SCC9 34.5 58.6

SW48 31.8 49.6

SW403 36.2 43

T84 13.6 14.6

colo205 2 4

dld1 1 3

H650 3 3

HCT8 5.5 1

kato3 2.5 3.5

LS1034 0 5.4

PC3 2.2 0

Correlation of MEDI3622 and cetuximab anti-proliferative activity. Indicated cell lines were incubated with 400 nM MEDI3622 or 200 nM cetuximab to determine maximum inhibition of each antibody as indicated in Materials and Methods. Percent inhibition was determined on day 4 using Cell Titer Glow. Results for each cell line are indicated.

Page 17: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

Supplementary Table 2fold decrease in protein shedding gene

7.817 NQO13.3399 PNPO2.6787 DPCD2.3452 RPUSD22.307 TPP12.0503 PTPRF2.0349 COG41.8526 PDXDC11.8002 SPINT11.7911 DLD1.7397 EXOSC101.7384 ACBD61.7291 PTK71.711 DAK1.6936 GPX21.6754 RBM31.6535 TXNRD11.6204 SEMA4D

Identification of ADAM17 substrates in COLO205 cells by SILAC proteomics. COLO205 cells at 75% confluence were preincubated with either 400 nM MEDI3622 or 400 nM nonspecific IgG. ADAM17 was then activated by addition of 50 nM PMA for 6 hours and cell supernatants were combined and analyzed by mass spectroscopy as described in Materials and Methods. Highlighted genes are shared with OE21 SILAC results.

Page 18: Supplementary Figure 1. Supplementary Figure 2 Supplementary Figure 3 A B

fold decrease in protein

shedding gene4.955401 PTK74.881859 DESP3.691127 AREG3.483835 GSN3.255738 MRC23.106941 MAP2K13.105494 GPC12.933756 PTPRG2.858531 PODXL22.807885 UBLA42.806151 PVRL12.778241 ZNF2922.683267 SEMA4C2.54868 PTPLAD1

2.539231 SEMA4D2.388973 MERTK2.314226 SORL12.222173 PRRC12.148228 MUC162.128973 PTPRS2.088991 XRN2

2.043945 PTPRF

fold decrease in protein

shedding gene2.024742 GPC42.005254 PWP21.986176 NRP11.972114 SDSL1.831032 SDC21.791858 CP1.787981 PHGDH1.738012 F11R1.716561 amigo21.713238 notch21.678106 HLA-A1.676502 ST141.671877 JaG11.668446 NES1.663395 MUC11.636233 DAG11.621587 GEG11.618359 LXN1.618044 VLDLR1.595405 LRP11.58985 ALCAM

1.568062 ULBP2

Supplementary Table 3

Identification of ADAM17 substrates in H358 lung cancer cells by SILAC proteomics. H358 cells at 75% confluence were preincubated with either 400 nM MEDI3622 or 400 nM nonspecific IgG. ADAM17 was then activated by addition of 50 nM PMA for 6 hours and cell supernatants were combined and analyzed by mass spectroscopy as described in Materials and Methods.


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