Fig. 4: Mannose induces PKM2 nuclear translocation by regulating K433 lactylation.

A MS identification of K433 on PKM2 as a potential lactylation site in bladder cancer cells. B Immunoprecipitation validation of lactylation of PKM2 identified by mass spectrometry. C Validation of the regulatory effects of mannose on the identified lactylation of PKM2 in bladder cancer cells. D Introduction of exogenous wild-type PKM2 and K433R mutant PKM2 in bladder cancer cells, comparing lactylation levels with those of lactate supplementation. E Comparison of pyruvate production by wild-type PKM2 and K433R mutant PKM2 in bladder cancer cells. F Immunoprecipitation validation of the interaction between PKM2 and p300 in bladder cancer cells. G Changes in PKM2 lactylation levels in bladder cancer cells after p300 was knocked down with two different siRNAs. H Effects of p300 knockdown on pyruvate kinase activity in bladder cancer cells. I Effects of mannose and lactate supplementation on the lactylation and acetylation levels of PKM2 in bladder cancer cells. J Validation of PKM2 phosphorylation in mannose-treated bladder cancer cells. K Fluorescence detection of PKM2 localization following mannose treatment. L Analysis of nuclear and cytoplasmic PKM2 expression in bladder cancer cells. M Examination of the effects of mannose, glucose, and lactate supplementation on nuclear PKM2 expression in bladder cancer cells.