Fig. 6

Synergistic effects of KTC1101 and anti-PD-1 therapy in B16 tumor models. A Schematic representation of various treatment strategies in B16 mouse models. (B-D) C57BL/6J mice with subcutaneous tumors of B16 cells were treated with either a vehicle, KTC1101, anti-PD-1 antibody, or KTC1101 and anti-PD-1 antibody for 2 weeks (n = 5). Images of the mice post-euthanasia and the excised tumors are presented (B). Tumor volumes (C) and body weights (D) were measured every two days. E Individual tumor growth curves of B16 tumor-bearing mice under different treatment conditions (n = 8). F Survival analysis of B16 tumor-bearing mice subjected to various treatments (n = 8). G Flow cytometric analysis of CD8⁺ and CD4⁺ T cell populations (gated on CD3⁺) in B16 tumors (n = 5). H Flow cytometric analysis of regulatory T cell (Treg cell; CD25⁺Foxp3⁺) populations in B16 tumors (n = 5). I Quantitative assessment of CD8⁺ T cell populations in B16 tumors (n = 5). J Quantification of CD4⁺ T cell populations in B16 tumors (n = 5). K Quantification of Treg cell populations in B16 tumors (n = 5). L Calculation of CD8⁺ T cells/Treg ratios in B16 tumors (n = 5). M Quantification of CD8⁺/CD4⁺ T cell ratios in B16 tumors (n = 5). N C57BL/6J mice with subcutaneous tumors of B16 cells were treated with either a vehicle or anti-PD-1, followed by RNA-Seq of the tumor tissues after 3 days. GSEA plots illustrating changes in “Interferon alpha response,” “Interferon-gamma response” pathways. O RT-PCR analysis of IFNγ expression levels in tumor tissues of a group of vehicle or anti-PD-1. P ELISA-based quantification of IFNγ secretion levels in tumor supernatants. Graphs are presented as the mean ± SEM from three independent experiments; p-values were determined using a two-tailed unpaired Student’s t-test; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001