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Supplementary Figure.1 NF- κ B p65 / β- actin protein ** ID8HM-1 1 2 3 ** * * * β-actin ID8 HM-1 +CPA(0.7uM)non-treat+CPA(7uM)+CPA(70uM) NF-κB p65 β-actin.

Published byJonah Welch Modified over 9 years ago

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Presentation on theme: "Supplementary Figure.1 NF- κ B p65 / β- actin protein ** ID8HM-1 1 2 3 ** * * * β-actin ID8 HM-1 +CPA(0.7uM)non-treat+CPA(7uM)+CPA(70uM) NF-κB p65 β-actin."— Presentation transcript:

1 Supplementary Figure.1 NF- κ B p65 / β- actin protein ** ID8HM-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 AB 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) * ** * * * *

2 Supplementary Figure.2 PD-L1 β-actin NF-κB p65 ID8 PD-L1 NF-κB p65 β-actin HM-1 +GEM(24uM)+PTX(2uM)non-treat+CBDCA(20uM)+GEM(120uM)+GEM(600uM)+PTX(10uM)+PTX(20uM)+CBDCA(100uM)+CBDCA(500uM) +GEM(24uM)+PTX(2uM)non-treat+CBDCA(20uM)+GEM(120uM)+GEM(600uM)+PTX(10uM)+PTX(20uM)+CBDCA(100uM)+CBDCA(500uM)

3 Supplementary Figure.3 NF-κB p65 β-actin +GEM+PTX+GEM+PTX ovary1847SK-OV-3 non-treat NF-κB p65 β-actin +GEM+PTX+GEM+PTX RMGIIOVCAR8 non-treat PD-L1 ovary1847 MHC class I β-actin +GEM(24uM)+PTX(2uM)non-treat+GEM(120uM)+GEM(600uM)+PTX(10uM)+PTX(20uM) β-actin +GEM(24uM)+PTX(2uM)non-treat+GEM(120uM)+GEM(600uM)+PTX(10uM)+PTX(20uM) PD-L1 SK-OV-3 MHC class I

4 PTXGEMCBDCAnon-treat ID8HM-1 PTXGEMCBDCAnon-treat 0 20 40 60 * ** * MHC class I protein Supplementary Figure.4 C ---- -+-+ +-+- ---- -+-+ +-+- OVCAR8 RMGII GEM PTX MHC class I β-actin MHC class I / β- actin protein RMGIIOVCAR8 non-treat PTX GEM PTX GEM non-treat 0.0 0.5 2.0 2.5 ** 1.0 1.5 A B MHC class I / β- actin protein SK-OV-3 ovary1847 0 1 2 3 ** ** +GEM(24uM) +PTX(2uM) non-treat +GEM(120uM)+GEM(600uM) +PTX(10uM)+PTX(20uM) +GEM(24uM) +PTX(2uM) non-treat +GEM(120uM)+GEM(600uM) +PTX(10uM)+PTX(20uM) ** **

5 Supplementary Figure. 5 P=0.002 DTX and CAPE PD-L1/GAPDH mRNA beforeafter 6.5 6.0 5.5 5.0

6 PTX GEM CBDCA non-treat PD-L1 protein ID8HM-1 PTX GEM CBDCA non-treat 0 5 10 15 ** PD-L1 / β- actin protein ID8HM-1 1 2 3 0 Supplementary Figure.6 A B ** * * PD-L1 β-actin PD-L1 β-actin ID8 HM-1 +IFN-γ(500pg/ml)non-treat+IFN-γ(1ng/ml)+IFN-γ(4ng/ml) PD-L1 β-actin ID8 HM-1 +CPA(0.7uM)non-treat+CPA(7uM)+CPA(70uM) PD-L1 β-actin CD +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) +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) * ** * * *

7 ---- -+-+ +-+- ---- -+-+ +-+- OVCAR8RMGII GEM PTX β-actin PD-L1 RMGIIOVCAR8 PD-L1 / β- actin protein 2.0 1.5 1.0 0.5 0.0 2.5 PTXGEMnon-treatPTXGEMnon-treat ** PD-L1 OVCAR8 13.2832.2127.2 14.4149.2971.26 RMGII 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 PD-L1 protein SK-OV-3 ovary 1847 RMGIIOVCAR8 PTXGEMnon-treatPTXGEMnon-treatPTXGEMnon-treatPTXGEMnon-treat 80 60 40 20 0 100 ** Supplementary Figure.7 A B C PD-L1 / β- actin protein 2 1 0 3 D +GEM(24uM) +PTX(2uM) non-treat +GEM(120uM)+GEM(600uM) +PTX(10uM)+PTX(20uM) +GEM(24uM) +PTX(2uM) non-treat +GEM(120uM)+GEM(600uM) +PTX(10uM)+PTX(20uM) SK-OV-3 ovary1847 ** ** **

8 Supplementary Figure. 8 NF-κB p65 β-actin ID8shRela69ID8shcControlID8shRela70HM-1shRela69HM-1shControlHM-1shRela70 B Rela/GAPDH mRNA shControl shRela69shRela70 ID8HM-1 shControl shRela69shRela70 A 3 2 1 0 4 C NF- κ B p65 / β- actin protein shControl shRela69shRela70 ID8HM-1 shControl shRela69shRela70 1.5 1.0 0.5 0.0 2.0 2.5 * * * * * ** *

9 MHC class I protein ID8shRela69 ID8shControl non-treat PTX GEM non-treat PTX GEM 40 20 0 80 60 * ** Supplementary Figure.9

10 SK-OV-3 ovary 1847 OVCAR8 NF- κ B p65 / β- actin protein siControlsiNF-κB p65siControlsiNF-κB p65siControlsiNF-κB p65 PTX non-treat GEM 2.0 1.5 1.0 0.5 0.0 2.5 A ** MHC class I GEM PTX ---- -+-+ +-+- ---- -+-+ +-+- OVCAR8 siControl OVCAR8 siNF-κB p65 β-actin NF-κB p65 PD-L1 Supplementary Figure.10 C OVCAR8 2.0 1.5 1.0 0.5 0.0 2.5 ** siControlsiNF-κB p65 GEM PTX non-treat MHC class I / β- actin protein GEM PTX non-treat PD-L1 / β- actin protein OVCAR8 2.0 1.5 1.0 0.5 0.0 2.5 ** siControlsiNF-κB p65 D B

11 PD-L1 NF-κB p65 β-actin +GEM+PTX+GEM+PTXsiControl siNF-κB p65 MHC class I ovary1847 Supplementary Figure.11 +GEM+PTX+GEM+PTXsiControl PD-L1 NF-κB p65 β-actin MHC class I SK-OV-3 siNF-κB p65 +GEM+PTX+GEM+PTXsiControl siNF-κB p65 PD-L1 NF-κB p65 β-actin MHC class I OVCAR8

12 Supplementary Figure.12 A NF- κ B p65 / β- actin protein siControlsiNF-kB p65 GEM PTX non-treat siControlsiNF-kB p65 ID8HM-1 1.5 1.0 0.5 0.0 2.0 2.5 ** NF-κB p65 GEM PTX ---- -+-+ +-+- ---- -+-+ +-+- HM-1 siNF-κB p65 HM-1 siControl 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- actin protein GEM non-treat PTX GEM non-treat PTX GEM non-treat PTX PD-L1 / β- actin protein shRela69 shRela70 shControl 2.0 1.5 1.0 0.5 0.0 2.5 ** GEM PTX ---- -+-+ +-+- ---- -+-+ +-+- HM-1 shRela69 HM-1 shRela70 β-actin PD-L1 ---- -+-+ +-+- HM-1 shControl C

13 PD-L1 NF-κB p65 β-actin +GEM+PTX+GEM+PTX ID8 siControl ID8 siNF-κB p65 NF-κB p65 β-actin +GEM+PTX+GEM+PTX HM-1 siControl HM-1 siNF-κB p65 PD-L1 Supplementary Figure.13 non-treat

14 PD-L1 β-actin +GEM+PTX+GEM+PTX ID8 shRela69 ID8 ShRela70 +GEM+PTX ID8 ShControl +GEM+PTX+GEM+PTX+GEM+PTX PD-L1 β-actin Supplementary Figure.14 non-treat HM-1 shRela69 HM-1 ShRela70 HM-1 ShControl

15 Supplementary Figure. 15 ID8 shControlID8 shIfngr1 IFNGR1 629.25 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4

16 PD-L1 NF-κB p65 β-actin Supplementary Figure. 16 IFN-γ GEM PTX ID8shIfngr1ID8shControl ------ --+--+ -+--+- +--+-- ------ --+--+ -+--+- +--+--

17 Supplementary Figure.17 controlGEM NF-κB p65 PD-L1 CD4 + TILs CD8 + TILs NF- κ B p65 staining score controlGEM 20 15 10 5 0 PD-L1staining score controlGEM 15 10 5 0 CD8 + T cell numbers/200 ╳ controlGEM 15 10 5 0 CD4 + T cell numbers/200 ╳ controlGEM 40 30 20 10 0 ** * * B %CD107a + cell PTX -pretreated ID8-control ID8-pdl1KO OVA loading 15 1010 5 0 * ** * -------- +-+++-++ ++-+++-+ -+---+-- A

18 Supplementary Figure. 18 B A ID8pdl1 PD-L1/GAPDH mRNA PTXGEM ID8Mirpdl1 non-treat 0.015 0.010 0.005 0.000 0.020 GEM PTX ---- -+-+ +-+- ---- -+-+ +-+- ID8MirControlID8pdl1 β-actin NF-κB p65 PD-L1 ---- -+-+ +-+- ID8Mirpdl1 PD-L1/GAPDH mRNA non-treatPTXGEM ID8MirControl 0.06 0.04 0.02 0.00 0.08 ** PD-L1/GAPDH mRNA non-treatPTXGEM 6 4 2 0 **

19 Supplementary figure.19 IgG IgG + GEM αPD-1 αPD-1 +GEM %survival days B 60 40 20 0 80 100 050100 A ID8-pdl1KO ID8-pdl1KO + GEM ID8-control + GEM ID8-control %survival days ID8-pdl1 ID8-pdl1 + GEM 60 40 20 0 80 100 050100

20 Supplementary Figure.20 C GEMPTXID8OVA PD-L1 β-actin NF-κB p65 OVA β-actin ID8 OVA 1ID8 OVA 2ID8 lacZ A B * %lysis E/T ration ID8OVA ID8lacZ with OVA loading ID8lacZ 150 100 50 0 131030 D PD-L1/GAPDH mRNA ID8lacZID8OVA GEM PTX non-treat 0.06 0.04 0.02 0.00 **

21 Supplementary Figure 21 A B CD4 + TILs/tumor(%) controlPTXαPD-1αPD-1+PTX 0.06 0.04 0.02 0.00 0.08 0.10 * * * CD8 + TILs/tumor(%) controlPTXαPD-1αPD-1+PTX 0.15 0.10 0.05 0.00 0.20 * * * C PD-1 + TILs/TILs(%) controlPTXαPD-1αPD-1+PTX 0.3 0.2 0.1 0.0 0.4 0.5 ** *

22 Supplementary Figure.22 controlPTX Gr-1 controlGEM Gr-1 Gr-1 + cell numbers/200 ╳ controlPTX 25 20 15 10 5 0 Gr-1 + cell numbers/200 ╳ controlGEM 25 20 15 10 5 0 *

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