Supplementary Figure.1 NF- κ B p65 / β- actin protein ** ID8HM-1 1 2 3 ** * * * β-actin ID8 HM-1...
22
Supplementary Figure.1 NF-κB p65 /β-actin protein ** ID8 HM-1 1 2 3 ** ** * * * β-actin ID8 HM-1 +CPA(0.7uM) non-treat +CPA(7uM) +CPA(70uM) NF-κB p65 β-actin NF-κB p65 A B 0 +GEM(24uM) +PTX(2uM) non-treat +CBDCA(20uM) +GEM(120uM) +GEM(600uM) +PTX(10uM) +PTX(20uM) +CBDCA(500uM) +CBDCA(100uM) +CPA(0.7uM) +CPA(7uM) +CPA(70uM) 0 +GEM(24uM) +PTX(2uM) non-treat +CBDCA(20uM) +GEM(120uM) +GEM(600uM) +PTX(10uM) +PTX(20uM) +CBDCA(500uM) +CBDCA(100uM) +CPA(0.7uM) +CPA(7uM) +CPA(70uM) * ** ** ** ** * * * * **
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Transcript of Supplementary Figure.1 NF- κ B p65 / β- actin protein ** ID8HM-1 1 2 3 ** * * * β-actin ID8 HM-1...
Supplementary Figure.1
NF-
κB
p6
5 /
β-a
ctin
pro
tein
**
ID8 HM-1
1
2
3
****
** *
β-actin
ID8
HM-1+
CPA
(0.7
uM
)
non-t
reat
+C
PA(7
uM
)
+C
PA(7
0uM
)
NF-κB p65
β-actinNF-κB p65
A B
0
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+C
BD
CA
(20
uM
)
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+C
BD
CA
(50
0uM
)+
CB
DC
A(1
00
uM
)
+C
PA(0
.7uM
)+
CPA
(7uM
)+
CPA
(70
uM
)
0
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+C
BD
CA
(20
uM
)
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+C
BD
CA
(50
0uM
)+
CB
DC
A(1
00
uM
)
+C
PA(0
.7uM
)+
CPA
(7uM
)+
CPA
(70
uM
)
*
**** **
*** **
*
**
Supplementary Figure.2
PD-L1
β-actin
NF-κB p65
ID8
PD-L1
NF-κB p65
β-actin
HM-1
+G
EM
(24
uM
)
+PTX
(2u
M)
non
-tre
at
+C
BD
CA
(20
uM
)
+G
EM
(12
0u
M)
+G
EM
(60
0u
M)
+PTX
(10
uM
)
+PTX
(20
uM
)
+C
BD
CA
(10
0u
M)
+C
BD
CA
(50
0u
M)
+G
EM
(24
uM
)
+PTX
(2u
M)
non
-tre
at
+C
BD
CA
(20
uM
)
+G
EM
(12
0u
M)
+G
EM
(60
0u
M)
+PTX
(10
uM
)
+PTX
(20
uM
)
+C
BD
CA
(10
0u
M)
+C
BD
CA
(50
0u
M)
Supplementary Figure.3
NF-κB p65
β-actin
+G
EM
+PTX
+G
EM
+PTX
ovary1847 SK-OV-3
non-t
reat
non-t
reat
NF-κB p65
β-actin
+G
EM
+PTX
+G
EM
+PTX
RMGII OVCAR8
non-t
reat
non-t
reat
PD-L1
ovary1847
MHC class I
β-actin
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)
+G
EM
(60
0uM
)
+PTX
(10
uM
)
+PTX
(20
uM
)
β-actin
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)
+G
EM
(60
0uM
)
+PTX
(10
uM
)
+PTX
(20
uM
)
PD-L1
SK-OV-3
MHC class I
PTX
GEM
CB
DC
A
non-t
reat
ID8 HM-1
PTX
GEM
CB
DC
A
non-t
reat0
20
40
60
*****
*
****
MH
C c
lass
I p
rote
in
Supplementary Figure.4
C
--
-+
+-
--
-+
+-
OVCAR8RMGIIGEMPTX
MHC class I
β-actin
MH
C c
lass
I /
β-a
ctin
pro
tein
RMGII OVCAR8
non-t
reat
PTX
GEM
PTX
GEM
non-t
reat0.0
0.5
2.0
2.5 ******
**
1.0
1.5
A B
MH
C c
lass
I /
β-a
ctin
pro
tein
SK-OV-3ovary1847
0
1
2
3
* *
**
**
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
* *
**
**
Supplementary Figure. 5
P=0.002
DTX and CAPE
PD
-L1
/GA
PD
H m
RN
A
before after
6.5
6.0
5.5
5.0
PTX
GEM
CB
DC
A
non-t
reat
PD
-L1
pro
tein
ID8 HM-1
PTX
GEM
CB
DC
A
non-t
reat0
5
10
15
**
****
**
****
PD
-L1
/β
-act
in p
rote
in ID8 HM-1
1
2
3
0
Supplementary Figure.6
A B
** **
*** *
PD-L1β-actinPD-L1β-actin
ID8
HM-1+
IFN
-γ(5
00
pg/m
l)
non-t
reat
+IF
N-
γ(1
ng/m
l)+
IFN
-γ(4
ng/m
l)
PD-L1
β-actin
ID8
HM-1
+C
PA(0
.7uM
)
non-t
reat
+C
PA(7
uM
)
+C
PA(7
0uM
)
PD-L1
β-actin
C D
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+C
BD
CA
(20
uM
)
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+C
BD
CA
(50
0uM
)+
CB
DC
A(1
00
uM
)
+C
PA(0
.7uM
)+
CPA
(7uM
)+
CPA
(70
uM
)
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+C
BD
CA
(20
uM
)
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+C
BD
CA
(50
0uM
)+
CB
DC
A(1
00
uM
)
+C
PA(0
.7uM
)+
CPA
(7uM
)+
CPA
(70
uM
)
* ** ** ** ***
**
**
** **
--
-+
+-
--
-+
+-
OVCAR8RMGII
GEMPTX
β-actin
PD-L1
RMGII OVCAR8
PD
-L1
/β
-act
in p
rote
in
2.01.51.00.50.0
2.5PTX
GEM
non-t
reat
PTX
GEM
non-t
reat
**
** ****
PD-L1
OVCAR813.28 32.21 27.2
14.41 49.29 71.26RMGII
100101102103104 100101102103104100101102103104
PD
-L1
pro
tein
SK-OV-3ovary1847 RMGII OVCAR8
PTX
GEM
non-t
reat
PTX
GEM
non-t
reat
PTX
GEM
non-t
reat
PTX
GEM
non-t
reat
80
60
40
20
0
100
**
**
**
****
******
Supplementary Figure.7
A B
C
PD
-L1
/β
-act
in p
rote
in
2
1
0
3
D
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
+G
EM
(24
uM
)
+PTX
(2uM
)
non-t
reat
+G
EM
(12
0uM
)+
GEM
(60
0uM
)
+PTX
(10
uM
)+
PTX
(20
uM
)
SK-OV-3ovary1847
* *
**
*** *
**
**
Supplementary Figure. 8
NF-κB p65
β-actin
ID8
shR
ela
69
ID8
shcC
ontr
ol
ID8
shR
ela
70
HM
-1sh
Rela
69
HM
-1sh
Contr
ol
HM
-1sh
Rela
70
B
Rela
/GA
PD
H m
RN
A
shC
ontr
ol
shR
ela
69
shR
ela
70
ID8 HM-1
shC
ontr
ol
shR
ela
69
shR
ela
70
A
3
2
1
0
4
C
NF-
κB
p6
5 /
β-a
ctin
pro
tein
shC
ontr
ol
shR
ela
69
shR
ela
70
ID8 HM-1
shC
ontr
ol
shR
ela
69
shR
ela
70
1.5
1.0
0.5
0.0
2.0
2.5
**
**
** *
*
MH
C c
lass
I p
rote
in
ID8shRela69
ID8shControl
non-t
reat
PTX
GEM
non-t
reat
PTX
GEM
40
20
0
80
60*
**
Supplementary Figure.9
SK-OV-3ovary1847 OVCAR8
NF-
κB
p6
5 /
β-a
ctin
pro
tein
siC
ontr
ol
siN
F-κ
B p
65
siC
ontr
ol
siN
F-κ
B p
65
siC
ontr
ol
siN
F-κ
B p
65
PTX non-treat
GEM
2.0
1.5
1.0
0.5
0.0
2.5
A
****
****
****
MHC class I
GEMPTX
--
-+
+-
--
-+
+-
OVCAR8siControl
OVCAR8siNF-κB p65
β-actin
NF-κB p65
PD-L1
Supplementary Figure.10
COVCAR8
2.0
1.5
1.0
0.5
0.0
2.5**
**
siControl siNF-κB p65
GEM PTX
non-treat
MH
C c
lass
I /
β-a
ctin
pro
tein
GEM PTX
non-treat
PD
-L1
/β
-act
in p
rote
in OVCAR8
2.0
1.5
1.0
0.5
0.0
2.5 ****
siControl siNF-κB p65
D
B
PD-L1
NF-κB p65
β-actin
+G
EM
+PTX
+G
EM
+PTX
siC
ontr
ol
siN
F-κ
B p
65
MHC class I
ovary1847
Supplementary Figure.11
+G
EM
+PTX
+G
EM
+PTX
siC
ontr
ol
PD-L1
NF-κB p65
β-actin
MHC class I
SK-OV-3
siN
F-κ
B p
65
+G
EM
+PTX
+G
EM
+PTX
siC
ontr
ol
siN
F-κ
B p
65
PD-L1
NF-κB p65
β-actin
MHC class I
OVCAR8
Supplementary Figure.12
A
NF-
κB
p6
5 /
β-a
ctin
pro
tein
siC
ontr
ol
siN
F-k
B p
65
GEM PTX
non-treat
siC
ontr
ol
siN
F-k
B p
65
ID8 HM-1
1.5
1.0
0.5
0.0
2.0
2.5
**
****
**
NF-κB p65
GEMPTX
--
-+
+-
--
-+
+-
HM-1siNF-κB p65
HM-1siControl
β-actin
PD-L1
B
1.0
0.5
0.0
2.0
1.5
siControlsiNF-κB p65
GEM PTX
non-treat
**
**
PD
-L1
/β
-act
in p
rote
in
GEM
non-t
reat
PTX
GEM
non-t
reat
PTX
GEM
non-t
reat
PTXP
D-L
1 /
β-a
ctin
pro
tein
shRela69shRela70shControl
2.01.51.00.50.0
2.5**
**
GEMPTX
--
-+
+-
--
-+
+-
HM-1shRela69
HM-1shRela70
β-actin
PD-L1
--
-+
+-
HM-1shControl
C
PD-L1
NF-κB p65
β-actin
+G
EM
+PTX
+G
EM
+PTX
ID8siControl
ID8siNF-κB p65
NF-κB p65
β-actin
+G
EM
+PTX
+G
EM
+PTX
HM-1siControl
HM-1siNF-κB p65
PD-L1
Supplementary Figure.13
non-t
reat
non-t
reat
non-t
reat
non-t
reat
PD-L1
β-actin
+G
EM
+PTX
+G
EM
+PTX
ID8shRela69
ID8ShRela70
+G
EM
+PTX
ID8ShControl
+G
EM
+PTX
+G
EM
+PTX
+G
EM
+PTX
PD-L1
β-actin
Supplementary Figure.14
non-t
reat
non-t
reat
non-t
reat
non-t
reat
non-t
reat
non-t
reat
HM-1shRela69
HM-1ShRela70
HM-1ShControl
Supplementary Figure. 15
ID8 shControl ID8 shIfngr1
IFNGR1
62 9.25
100101102103104100101102103104
PD-L1
NF-κB p65
β-actin
Supplementary Figure. 16
IFN-γGEMPTX
ID8shIfngr1ID8shControl
---
--+
-+-
+--
---
--+
-+-
+--
Supplementary Figure.17
control GEM
NF-κB p65
PD-L1
CD4+ TILs
CD8+ TILsN
F-κB
p6
5
stain
ing
sco
re
control
GEM
20151050
PD
-L1
st
ain
ing
sco
re
control GEM
15
10
5
0
CD
8+ T
cell
nu
mb
ers
/20
0╳
control GEM
15
10
5
0
CD
4+ T
cell
nu
mb
ers
/20
0╳
control GEM
403020100
**
**
*
*
B
%C
D1
07
a+ce
ll
PTX -pretreatedID8-controlID8-pdl1KO
OVA loading
15
10
5
0
***
*
----
+-++
++-+
-+--
A
Supplementary Figure. 18
B
AID8pdl1
PD
-L1
/GA
PD
H m
RN
A
PTX GEM
ID8Mirpdl1
non-treat
0.015
0.010
0.005
0.000
0.020
GEMPTX
--
-+
+-
--
-+
+-
ID8MirControl
ID8pdl1
β-actin
NF-κB p65
PD-L1
--
-+
+-
ID8Mirpdl1
PD
-L1
/GA
PD
H m
RN
A
non-treat
PTX GEM
ID8MirControl
0.06
0.04
0.02
0.00
0.08
****
PD
-L1
/GA
PD
H m
RN
A
non-treat PTX GEM
6
4
2
0
****
Supplementary figure.19
IgGIgG + GEMαPD-1 αPD-1 +GEM
%su
rviv
al
days
B
60
40
20
0
80
100
0 50 100
A
ID8-pdl1KOID8-pdl1KO + GEM
ID8-control + GEMID8-control
%su
rviv
al
days
ID8-pdl1ID8-pdl1 + GEM
60
40
20
0
80
100
0 50 100
Supplementary Figure.20
C
GEM
PTX
ID8
OVA
PD-L1
β-actin
NF-κB p65
OVA
β-actin
ID8
OVA
1
ID8
OVA
2
ID8
lacZ
A B
*%ly
sis
E/T ration
ID8OVA ID8lacZ with OVA loading
ID8lacZ 150
100
50
0131030
D
PD
-L1
/GA
PD
H m
RN
A
ID8lacZ ID8OVA
GEM PTX
non-treat 0.06
0.04
0.02
0.00
****
****
Supplementary Figure 21
A B
CD
4+ T
ILs/
tum
or(
%)
contr
ol
PTX
αPD
-1
αPD
-1+
PTX
0.060.040.020.00
0.080.10
**
*C
D8+
TIL
s/tu
mor(
%)
contr
ol
PTX
αPD
-1
αPD
-1+
PTX
0.15
0.10
0.05
0.00
0.20
**
*
C
PD
-1+
TIL
s/TIL
s(%
)
contr
ol
PTX
αPD
-1
αPD
-1+
PTX
0.30.20.10.0
0.40.5 **
*
Supplementary Figure.22
control PTX
Gr-1
control GEM
Gr-1
Gr-
1+ c
ell
nu
mb
ers
/20
0╳
control PTX
2520151050
Gr-
1+ c
ell
nu
mb
ers
/20
0╳
control GEM
2520151050
*
![Structure of the specificity domain of the Dorsal ... · NF-κB p50 [22,23], p52 [24] and p65 [25] have been reported. The crystal structure of the mouse p50/p65 het-erodimer bound](https://static.fdocument.org/doc/165x107/5e312dda7e32fa57ce774aa6/structure-of-the-specificity-domain-of-the-dorsal-nf-b-p50-2223-p52-24.jpg)
