Fig. 2

Optimized dose tucidinostat promotes CD8⁺ T cell migration by inducing pro-inflammatory CCL5 secretion in tumor. a Volcano plot showing the significantly overexpressed genes (red) and significant underexpressed genes (green) in tumor from CT26 tumor-bearing mice on day 10 post-treatment with tucidinostat (25 mg/kg, gavage, daily, n=3) or DMSO as vehicle control (DMSO, n=3). b Heatmap showing the scores of immune gene signatures in tumor from CT26 tumor-bearing mice on day 10 post-treatment initiation. Colors in the heatmap represent the level of significance of the enrichment (−log10 of the adjusted p values). c Relative mRNA expression of CCL5, CXCL9, CXCL10, and PD-L1 compared to vehicle (set to fold change = 1) in 4T1, LLC, and CT26 cells that were exposed to increasing concentrations (2.5, 5, 7.5 μM) of tucidinostat for 24 h. The experiment was performed in triplicate. d Relative protein expression of CCL5 and phospho-p65-NF-κB (pNF-κB p65) compared to vehicle in CT26 cells that were exposed to increasing concentrations (2.5, 5, 7.5 μM) of tucidinostat for 48 h. The experiment was performed in triplicate. e Relative protein expression of CCL5 and pNF-κB p65 compared to vehicle in CT26 cells that were exposed to tucidinostat and NF-κB inhibitor (BAY11-7082) for 48 h. The experiment was performed in triplicate. f Kaplan-Meier survival curves for overall survival and disease-free survival in three solid tumor types as stratified by CCL5^HighCD8A^High or CCL5^lowCD8A^low expression status using the TCGA database. g CD8⁺ T cell migration with different doses of CCL5 protein (10, 20, 50μg/ml) for 24h. The experiment was performed in triplicate. The error bars indicate mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 by one-way ANOVA. ns not significant, BRCA breast invasive carcinoma, LUAD lung adenocarcinoma, COAD colon adenocarcinoma, CON control group, Tuc tucidinostat, phospho-p65-NF-κB pNF-κB p65