Fig. 5: HDAC8-dependent deacetylation at K62 promotes the binding with β-catenin and upregulating cyclin D1 expression.

A HepG2 cells were co-transfected with HA-β-catenin with Flag-HADC8 (Flag-vector), after 48 h, cells were collected for immunoprecipitated by HA beads, analyzed by western blotting with PKM2 and HA antibodies. B Co-transfected HA-β-catenin and Flag-PKM2 plasmids for 24 h, then PCI-34051 (25 µM, DMSO as control) added for another 24 h, immunoprecipitated by HA beads, western blotting using Flag and HA antibodies. C Cells were transfected by PKM2 WT or K62R plasmids, separately. After 48 h, the Duollink PLA technology was used to illustrate the interaction (red) of HA-PKM2and β-catenin in HepG2 cells (Scale bar, 50 µm). D HepG2 were transfected with siRNA against HDAC8 or PKM2 (negative control siRNA) for western blotting or qPCR. Another half were analyzed by cytometry. E, F Based on which siRNA were transfected, after 6 h, the cells were overexpressed another protein expression plasmid (controlled by negative siRNA and vector plasmid). Then experiments were carried out as mentioned above. G Cells were transfected PKM2 WT or mutant plasmids, separately. After 48 h, lysates of HepG2 were analyzed by western blotting using the indicated antibodies. H Based on above result. cells were transfected PKM2 wild type or K62 mutant plasmids, separately, then PCI-34051 (25 µM) was added after 24 h and maintained for 24 h. Cell lysates were analyzed by western blotting using the indicated antibodies. (one way ANOVA in (D–F), *p < 0.05, **p < 0.01).