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FIGURE 1A
p38
EVSh
MAPK14
HCT116-TP53KO
GAPDH
EV EVSh
MAPK14bis
ShMAPK
12
ShMAPK
13EV
ShMAPK
11
B
0.0
0.5
1.0
1.5
2.0
2.5
ShLUC
ShMAPK
14
ShMAPK14bis
ShMAPK
11
ShMAPK
12
ShMAPK
13
IC50
(nM
)
HCT116-TP53KO
HA
MA
PK
14 C
A
MA
PK
11 C
A
MA
PK
12 C
A
MA
PK
13C
A
tubulin
HCT116-TP53KO
D Kinase Assay
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
EVMAPK
14CA
MAPK11CA
MAPK12CA
MAPK13CA
IC50
(nM
)
HCT116-TP53KO
Epp38
GAPDH
HCT116-TP53KO
NT SN38F
C
p ATF2
MA
PK
14 C
A
MA
PK
11 C
A
MA
PK
12 C
A
MA
PK
13C
A
FIGURE 2A
0
1
2
3
4
0 2 4 6 8 10 12
HCT116-TP53KO EV
HCT116-TP53KO-MAPK14CAC
ell n
umbe
r (1
06 )
Days
B
Dou
blin
g T
ime
(hou
rs)
020406080
100120
EV MAPK14-CA
HCT116-TP53KO
HCT116-TP53KO-EV
HCT116-TP53KO-MAPK14CA
0
20
40
60
80
100
120
D7 D14 D21 D28
% li
vin
g c
ells
CHCT116-TP53 KO
0
5
10
15
20
EVMAPK14
CA
% A
PO
PT
OS
IS
D
EHCT116-TP53KO-EV HCT116-TP53KO-MAPK14CA
0
1
2
3
4
5
HCT116-TP53KO
-EV
HCT116-TP53KO-
MAPK14CA
Blu
e ce
ll nu
mbe
r/fie
ld
FIGURE 3
AHCT116-TP53KO-EV HCT116-TP53KO-MAPK14CA
S49.4%
G0G1
28.8%
G2/M18.4%
S37.1%
G0G1
36.8%
G2/M19.3%
B
0
20
40
60
80
100
120
0 5 10 15 20 25 30
HCT116-TP53KO-EV
HCT116-TP53KO-MAPK14CA
% o
f Brd
U s
tain
ed c
ells
D
0.0
0.2
0.4
0.6
0.8
1.0
1.2
EVD7 D14 D21 D28
p21
Arb
itrar
y R
NA
uni
ts
MAPK14CA
HCT116-TP53KO
C
p21
Actin
EVD7 D14 D21 D28
MAPK14CA
HCT116-TP53KO
HA
p21
shLUC shp21
HCT116-TP53KO-MAPK14CA
GAPDH
E
HCT116-TP53KO-ShLuc-MAPK14CA
HCT116-TP53KO-Shp21-MAPK14CA
S25.8%
G0G1
45.6%
G2/M23.16%
S46.1%
G0G1
20.9%G2/M28.9%
F G
0102030405060708090
0 5 10 15 20 25 30
HCT116-TP53KO-ShLuc-MAPK14CA
HCT116-TP53KO-Shp21-MAPK14CA
% o
f Brd
U s
tain
ed c
ells
FIGURE 4A
HCT116-TP53KO
EV MAPK14CALC3
Actin
EVD7 D14 D21 D28
MAPK14CA
HCT116-TP53KO
H1299
LC3
Tubulin
EV D7 D14 D21
MAPK14CA
B
C
500 nm500 nm
HCT116-TP53KO
EV MAPK14CA
Aut
op
ha
goso
me
nu
mb
er/
cell
MAPK14CAEV
HCT116-TP53 KO
0123456
D
HCT116-TP53 KO
MAPK14CAEV
GFP-LC3 labelling
10-
8 -
6 -
4 -
2 -
0 -
EHCT116-TP53KO-
MAPK14CA
GAPDH
LC3
shLUC shp21
HCT116-TP53KO
GF
P-L
C3
spot
s/ce
ll
EV MAPK14CA
FIGURE 5
p62
Actin
EV D7 D14 D21 D28
MAPK14CA
HCT116-TP53KOA
0
1
2
EV D7 D14 D21 D28
p62
leve
l(a
rbitr
ary
unit)
MAPK14CA
HCT116-TP53KO
B HCT116-TP53KO (J14)
p62
NT 2H 4H 8H NT 2H 4H 8H
EV MAPK14CA
GAPDH
0
40
80
120
160
nt 2h 4h 8h nt 2h 4h 8h
p62
leve
l(a
rbitr
ary
unit)
EV
HCT116-TP53KO
MAPK14CA
NT 3MA Baf NT Baf 3MA
LC3
Actin
EV
HCT116-TP53KO
MAPK14CA D14
HCT116-TP53KO
EV MAPK14CA D14
0
0.5
1
1.5
2
2.5
3
NT 3MA Baf NT 3MA Baf
LC3-
II le
vel
(arb
itrar
y un
it)C
D E
Atg7
Actin
HCT116-TP53KO
Atg5
siCT D2 D4
siRNA
NT 3MA Baf NT 3MA Baf
HCT116-TP53KO
EV MAPK14CA D14
Cel
l via
bilit
y (%
)
0
20
40
60
80
100
HCT116-TP53KO
EV MAPK14CA D14
Cel
l via
bilit
y (%
)
0
20
40
60
80
100
siC
T
siA
tg 5
siA
tg 7
siC
T
siA
tg 5
siA
tg 7
FIGURE 6
A LC3 labelling
HCT116-TP53 KO
SN38
NT
LC3
spot
s/ce
ll
012
34
56
7
NT SN38
HCT116-TP53KO
B
NT SN
HCT116TP53KO
LC3
Actin
NT SN
SW480
LC3
Actin
C
SN38SN+
Baf
SN+
3MA
LC3
GAPDH
HCT116-TP53KO
D
siCT siAtg5 siAtg7
SW480
SN
38 IC
50 (
nM)
0
2
4
6
8 Cytotoxic Assay
E F
siCT siAtg5 siAtg7
4
3
2
1
0
HCT116-TP53KO
SN
38 IC
50 (
nM) Cytotoxic Assay
0
0.4
0.8
1.2
1.6
siCT siATG5 siATG7
HCT116-TP53KO
SN
38 IC
50 (
nM) Clonogenic Assay
HCT116-TP53KO
0
20
40
60
80
100
120
NT SN38 SN38+3MA
% li
ving
cel
ls
G
FIGURE 7
LC3
MAPK14
Actin
NT SN SN+3MA NT SN SN+3MA
HCT116-TP53KO-shLUC
HCT116-TP53KO-shMAPK14A
B
shLUC shMAPK
11
shMAPK
12
shMAPK
13
shLUC shMAPK
11
shMAPK
12
shMAPK
13
LC3
HCT116-TP53KO
GAPDH
NT SN38
0
1
2
3
HCT116-TP53KOShLuc
HCT116-TP53KOShMAPK14
SN38SN38+3MA
SN
38 IC
50 (
nM)C
D
siCT
siATG5
siATG7
0
1
2
3
HCT116-TP53KO ShLuc
SN
38 IC
50 (
nM)
HCT116-TP53KOShMAPK14
