Fig. 4

In vivo evaluation of KTC1101's therapeutic efficacy and immune modulation. A Schematic representation of treatment regimens applied in thestudy. B-D C57BL/6J mice with subcutaneous B16 cell tumors received either vehicle or KTC1101 treatments for 2 weeks (n = 5). Post-euthanasia images of the excised tumors (B), alongside measurements of tumor volumes (C) and body weights (D) every two days, are depicted. E Individual tumor growth tracking in B16 tumor-bearing mice subjected to KTC1101 or vehicle treatment (n = 8). F Survival curve analysis of B16 tumor-bearing mice under KTC1101 or vehicle treatment (n = 8). G C57BL/6J mice with subcutaneous tumors of B16 cells were treated with either a vehicle or KTC1101 for 3 days, followed by RNA-Seq of the tumor tissues. Scatter plot showcasing Gene Set Enrichment Analysis (GSEA) results for differentially expressed genes mapped to Gene Ontology (GO) pathways, including normalized enrichment scores and p-values, with horizontal dashed lines indicating p-values of 0.1, 0.05, 0.001, and 0.0001. H Ridgeline plot visualization of GSEA data highlighting the enrichment of specific pathways in response to KTC1101 treatment, including term names, p-values, and enrichment scores, with normalized enrichment scores represented as color variations. I Bubble plot displaying the results of Gene Set Variation Analysis (GSVA) for differentially expressed genes, with enrichment scores depicted as color variations and bubble size. J Box plot presenting the results of CIBERSORT analysis using 511 mouse gene signatures as a reference, including relative scores of immune cell types and cell population categories. K Representative flow cytometric analysis of CD8⁺ and CD4⁺ T cell populations (gated on CD3⁺) in B16 tumors (n = 5). L Representative flow cytometric analysis of regulatory T cell (Treg cell; CD25⁺Foxp3⁺) cell populations in B16 tumors (n = 5). M Quantification of CD8⁺ T cell populations in B16 tumors (n = 5). N Quantification of CD4⁺ T cell populations in B16 tumors (n = 5). O Quantification of Treg cell populations in B16 tumors (n = 5). P Quantification of CD8⁺ T cells/Treg ratios in B16 tumors (n = 5). Q Quantification of CD8⁺/CD4⁺ T cell ratios in B16 tumors (n = 5). 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