Fig 2: BXCL701 treatment enhances anti-PD1 efficacy in mT3-2D tumors. (A) Schematic representation of in vivo experimental design testing combination treatment of BXCL701 and anti-PD1 vs single agents alone and PBS controls in mT3-2D tumors. (B) DPP activity measured in the serum of PBS, anti-PD1, BXCL701 or anti-PD1 and BXCL701 (‘Combo’) treated mice (n=5 per group, each dot representing pooled serum from three mice, **p<0.01, ***p<0.001, ****p<0.0001 as determined by ANOVA followed by Tukey’s multiple comparison’s test, any pairwise comparisons not shown were not significant). (C) DPP activity measured in the tumor lysates of PBS, anti-PD1, BXCL701 and combo-treated mice (n=3 per group,*p<0,05 as determined by ANOVA followed by Tukey’s multiple comparison’s test, any pairwise comparisons not shown were not significant). (D) Average mT3-2D tumor growth curves in C57BL/6 mice (n=15 per group) treated with PBS, anti-PD1, BXCL701 and combo. Tumor growth was monitored weekly. (E) Tumor volumes for PBS, anti-PD1, BXCL701 and combo-treated mT3-2D tumors at the end of treatment (week 4). (***p<0.001, ns=nonsignificant as determined by ANOVA followed by Tukey’s multiple comparison’s test). (F) Flow cytometry data representing CD4+, CD8+ and NK1.1+ cells as percent of live cells in tumors collected from PBS andcombo-treated mice (n=3 per group, each dot representing three to four tumors from individual mice, *p<0.05, **p<0.01, as determined by unpaired two-tailed t-test). (G) Flow cytometry data representing percentage of live cells that are CXCR3+CD4+, CXCR3+CD8+ and CXCR3+NK1.1+ cells in tumors from PBS, anti-PD-1, BXCL701 and combo treated mice (n=3 per group, each dot representing three to four tumors from individual mice, *p<0.05, **p<0.01, ***p<0.001, ns=nonsignificant as determined by one-way ANOVA followed by Tukey’s multiple comparison test). (H) Flow cytometry data representing percentage of live cells that are CD4 +IFN?+Tbet+ Th1 cells in tumors from PBS, anti-PD-1, BXCL701 and combo treated mice (n=3 per group, each dot representing three to four tumors from individual mice, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns=nonsignificant as determined by one-way ANOVA followed by Tukey’s multiple comparison test). ANOVA, analysis of variance; DPP, dipeptidyl peptidase; PBS, phosphate-buffered saline.