Fig. 3: Enhanced H3K18la by glycolysis-derived lactate promotes the expression of Cxcl1.

A Lactylation levels of pan-lysine and different lysine sites in histone H3 of 7 pairs of PDAC tissue and adjacent normal tissue were detected by western blot. B Representative IHC images of clinical PDAC and adjacent normal tissue stained by H3K18la antibody (n = 79 patients). C The diagram illustrating the targets of 2DG, shRNA, and rotenone in the glycolysis process. D, E Relative levels of H3K18la and PanKla in PDAC cells treated with gradient doses of 2DG (D) or rotenone (E) for 24 h were detected by western blot. F Relative levels of H3K18la and PanKla in subcutaneous tumors treated with or without 2DG were detected by western blot. (n = 4 biologically independent samples per group). G H3K18la levels of subcutaneous tumors treated with or without 2DG were detected by immunofluorescence staining. H The H3K18la levels in PDAC cells treated with 2DG or sodium lactate were detected by western blot. I The H3K18la level in PANC1 and PANC02 cells following LDH-knockdown and sodium lactate treatment by western blot. J Schematic diagram of the CUT&Tag assay for PanKla or H3K18la in BxPC3 cells with or without 2DG treatment. K TSS heatmaps presenting the binding density of PanKla and H3K18la with or without 2DG treatment. L Venn diagram indicating the overlap of differentially expressed genes in TCGA-PAAD between PDAC and adjacent normal tissue, differentially expressed genes in RNA-seq after 2DG treatment, and differentially bound genes of H3K18la and PanKla after 2DG treatment. M Genome browser track analysis shows the PanKla and H3K18la levels in the Cxcl1 promoter region with or without 2DG treatments. N The Cxcl1 promoters that are bound by H3K18la and PanKla in PDAC cells were quantified using ChIP-qPCR assays (n = 3 independent experiments). O Relative mRNA levels of Cxcl1 in PDAC cells treated with 2DG or sodium lactate were detected by qRT-PCR (n = 3 independent experiments). P Representative IHC images stained by CXCL1 or H3K18la antibody and correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples (n = 21 patients). Scale bar = 100 μm. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test (N–O) and two-tailed Pearson’s correlation analysis (P). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results, where Histone H3, H3K9la, H3K14la, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. Source data are provided as a Source Data file.