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Counts TLR3-PE 100 80 60 40 20 0 Counts TLR8-PE 100 80 60 40 20 0 Counts TLR9-PE 100 80 60 40 20 0 C AB Supplemental Figure 1

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A 250 200 150 100 50 0 TLR7-PE No. of living cells (in million) B 1.5 1 0.5 0 PBS CL264 Cisplatin CL264+ Cisplatin Percentage (apoptotic cells) 50 40 30 20 10 0 PBS CL264 Cisplatin CL264+ Cisplatin Counts ** *** Supplemental Figure 2

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6000 4000 2000 0 14000 12000 10000 8000 PBS CL264 Cisplatin(100µM)Cisplatin(200µM) Cisplatin(300µM) CL264+Cisplatin(100µM)CL264+Cisplatin(200µM) CL264+Cisplatin(300µM) ns Tumor volume (mm 3 ) ** Supplemental Figure 3

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A B Tumor volume (mm 3 ) day 28 7500 6000 4500 3000 1500 0 CL264 Cisplatin IRS661 ODN -+-++-+ --++-++ ----+++ ++++--- - - + - Supplemental Figure 4

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10 20 40 30 0 PBS CL264 %CD3+ cells among CD45+ cells ns PBS CL264 0 5 10 15 20 nb CD3+ cells (x10 6 ) ns PBS CL264 %CD3+ CD4+ cells among CD45+ cells 80 60 100 40 20 0 ns PBS CL264 nb CD3+ CD4+ cells (x10 6 ) 0 5 10 15 ns CD3 + T cellsCD4 + T cells PBS CL264 2 4 8 6 0 10 %CD3+ CD8+ cells among CD45+ cells ns PBS CL264 nb CD3+ CD8+ cells (x10 6 ) 0 1 2 3 ns CD8 + T cells PBS CL264 0 2 4 6 8 %CD3+ Foxp3+ cells among CD45+ cells ns nb CD3+ Foxp3+ cells (x10 6 ) PBS CL264 0 1 2 3 ns CD3 + Foxp3 + T Reg cells PBS CL264 0 5 10 15 20 %Gr1- CD11c+ cells among CD45+ cells nb Gr1- CD11c+ cells (x10 6 ) 0 5 10 15 PBS CL264 Gr1 - CD11c + dendritic cells PBS CL264 %Gr1- CD11b+ cells among CD45+ cells 0 20 40 60 PBS CL264 0 0.5 1 1.5 nb Gr1- CD11b+ cells (x10 6 ) Gr1 - CD11b + macrophages PBS CL264 10 20 40 30 %Gr1+ CD11b+ cells among CD45+ cells 0 PBS CL264 0.5 1 1.5 0 nb Gr1+ CD11b+ cells (x10 6 ) Gr1 + CD11b + MDSCs A B CD EF G ** Supplemental Figure 5

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P value Percentage of TLR7 expressing tumor cells Supplemental Figure 6

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P value Percentage of TLR7 expressing tumor cells Supplemental Figure 7

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Overall Survival (OS) Time (months) 1.6.8.4 0.2 0 20 40 60 80 100 120 Responders =55/166 Non-responders =111/166 P = 0.00218 Supplemental Figure 8

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R 2 = 0.5084 p= 0.0009 % TLR7 in tumor cells in LN after chemotherapy % TLR7 in tumor cells in lung after chemotherapy 100 80 60 40 20 0 40 6080100 0 Supplemental Figure 9

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No Yes % TLR7 positive tumoral cells Tumor Downstaging A 100 80 60 40 20 0 B Yes 100 80 60 40 20 0 No Lymph Node Downstaging % TLR7 positive tumoral cells PatientsMean %TLR7 expressing cells P value for the mean Median %TLR7 expressing cells P value for the median Tumor downstaging Yes27 0.0053 0 0.0013 No4750 Lymph Node downstaging Yes28 0.0002 2 0.0003 No51 50 C Supplemental Figure 10

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