Fig. 5: H3K18la enrichment at FOXO3 and CXCL16 promoters upregulates CXCL16 expression in CAFs.

a IGV tracks presenting H3K18la enrichment at the CXCL16 gene locus in CAFs from MPE by CUT&Tag analysis. b ChIP–qPCR validation of H3K18la binding at the CXCL16 promoter. c–e Predicted FOXO3 binding sites at the CXCL16 promoter using JASPAR (http://jaspar.genereg.net). f ChIP–qPCR validation of FOXO3 binding at the CXCL16 gene promoter in CAFs. g Western blot analysis of FOXO3 and CXCL16 expression following FOXO3 knockdown or overexpression in CAFs. h, i CXCL16 levels in CAFs supernatants after FOXO3 knockdown (h) or overexpression (i). j Schematic of dual-luciferase reporter assay, drawn by Figdraw. k Luciferase assays assessing FOXO3-mediated regulation of CXCL16 transcription in CAFs. l, m CUT&Tag (l) and ChIP–qPCR (m) analyses showing H3K18la enrichment at the FOXO3 promoter. n Western blot analysis of the indicated proteins in CAFs treated with glucose (0–20 mm/l), DCA (0–20 mm/l), oxamate (0–20 mm/l), retenone (0–50 nm/l) or β-alanine (0–20 mm/l) for 48 h. o Western blot analysis of the indicated proteins in LDHA-knockdown CAFs. Data shown in b, f–i, k and m–o are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t-test (h and i) or one-way ANOVA (b, f, k and m). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns not significant, IGV Integrative Genomics Viewer, OE overexpression.