Fig. 6

Accumulated lactate boosts transcriptional activation of cell cycle-related gene CCNB1 by histone lactylation. A, B Cell cycle assessed by flow cytometry assays in indicated cells. C The results of qPCR indicating the mRNA of CCNB1, CDK1, RAD21 in indicated cells. D, E Histone of indicated cells were extracted and the level of H4K12la was detected by western blotting. The level of Histone H3 serves as control. F ChIP assay to detect histone lactylation level in CCNB1 promoters in PC9-BrM3 cells. (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, group #1, PC9 transfected with negative control plasmid; group #2, PC9 transfected with AKR1B10 plasmid; group #3, PC9 transfected with negative control plasmid and treated with sodium lactate 50 mM for 24 h; group #4, PC9-BrM3 cells transfected with negative control shRNA; group #5, PC9-BrM3 cells transfected with AKR1B10-targeted shRNA vector; group #6, PC9-BrM3 cells transfected with negative control shRNA and treated with GSK2837808A 10µM for 24 h)