Fig. 3: Hypoxia-induced lactate production upregulates CXCL16 expression in CAFs.

a GSVA of signaling pathways enriched in CXCL16⁺ CAFs compared with CXCL16⁻ CAFs. b, c Gene set enrichment analysis (GSEA) was performed on gene sets related to the hypoxia signaling pathway (b) and the glycolysis signaling pathway (c). A positive NES indicates higher expression in CXCL16⁺ CAFs. d, e Lactate levels in CAFs cultured under normoxia (21% oxygen) or hypoxia (1% oxygen) for 48 h or at indicated time points (n = 3). f, g CXCL16 concentrations in CAFs supernatant under normoxia or hypoxia for 48 h or at indicated time points (n = 3). h Western blot of CXCL16 expression in CAFs under hypoxia at designated times. i Schematic of metabolic modulators targeting glycolysis or lactate production. j, k Lactate levels (j) and CXCL16 concentrations (k) in CAFs treated with indicated glycolysis modulators for 48 h (n = 3). l–n Dose-dependent effects of DCA (l), oxamate (m) or rotenone (n) on lactate and CXCL16 levels in CAFs supernatant. o Western blot analysis of CXCL16 expression in CAFs treated with the indicated concentrations of glycolysis modulators for 48 h. Data shown in d–h and j–o are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t-test (d and f) or one-way ANOVA (e, g and j–n). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns not significant, NES normalized enrichment score, DMSO dimethylsulfoxide, DCA dichloroacetate.