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. 2021 May 20;12(6):517. doi: 10.1038/s41419-021-03782-w

Fig. 6. TSP50 acetylated PKM2 at K433 site.

Fig. 6

A Vectors encoding for multiple mutant PKM2 were constructed for subsequent analysis using a site-directed mutagenesis kit. B PKM2 K62R, K305R or K433R mutant vector was co-transfected with TSP50 into Huh7 cells. The cell was harvested and followed by IP with anti-Flag and PKM2 activity analysis. C–E The pcDNA3.1-TSP50 and Flag-PKM2 WT, Flag-PKM2K62R, Flag-PKM2 K305R or Flag-PKM2 K433R mutant plasmids were co-transfected into cells, respectively. The level of PKM2 K433 acetylation was detected using an anti-PKM2 K433ac antibody. F–R Acetylated mimic of PKM2 (PKM2-K433Q) transfected in cell lines deficient of TSP50, the PKM2 activity, cell viability, BrdU absorbance, glucose consumption, lactate production and ATP levels were detected. All aerobic glycolysis values were normalized to protein levels. N = 3 biologically independent replicates. t-Test statistical analysis was used. Data were presented as means ± s.d. *P < 0.05, **P < 0.01. ns, no significance.