Fig. 2: Deacetylation of PGAM2 at K100 promotes its ubiquitination and degradation.

A Half-life of PGAM2 was analyzed by western blot in HCC cell lines treated with 200 μg/mL cycloheximide (CHX) for indicated hours. B The curves were completed based on the gray value quantification of the bands in (A). C PGAM2 was measured by western blot in HCC cell lines treated with 25 μM proteasome inhibitor MG132 or 50 μM lysosome inhibitor Chloroquine for 16 h. D Construction and western blot analysis of HEK293T and HCCLM3 cell lines expressing PGAM2 wild type (PGAM2-WT) or deacetylation-mimic (PGAM2-K100R) proteins. E Acetylation levels of PGAM2-WT and PGAM2-K100R were detected by Co-IP. F, G Half-life of PGAM2-WT and PGAM2-K100R were detected by western blot in HEK293T and HCCLM3 cell lines treated with 200 μg/mL CHX. H Exogenous PGAM2-WT, PGAM2-K100R, and ubiquitin-expressing plasmids were transfected into HEK293T cells, then ubiquitination levels of PGAM2-WT and PGAM2-K100R were measured by Co-IP after being treated with 25 μM MG132 for 16 h. I, J Cell growth rates and colony formation abilities detection of PGAM2-WT and PGAM2-K100R expressing HCC cell lines. *p < 0.05, **p < 0.01, ***p < 0.001.