Fig. 3: SIRT2 deacetylates PGAM2 to promote HCC progression.

A Plasmids expressing Flag-PGAM2 and SIRT2 were transfected into HEK293T cells and lysates were co-immunoprecipitated with FLAG or SIRT2 antibodies and then immunoblotted with indicated antibodies. B Acetylation level of PGAM2 was detected by Co-IP in HEK293T cell line treated with 40 mM SIRTs family inhibitor Nicotinamide (NAM) or 20 μM HDAC Ι/П family inhibitor Trichostatin A (TSA) for 12 h. C Plasmids expressing PGAM2-WT and PGAM2-K100R were transfected into HEK293T cells with or without SIRT2-expressing plasmid, then acetylation levels of PGAM2-WT and PGAM2-K100R were detected by Co-IP. D Schematic of the domain structure of PGAM2 and SIRT2. E Interaction between SIRT2 and PGAM2 was analyzed by pymol, PGAM2 (red), SIRT2 (blue). F Plasmid expressing PGAM2-WT or PGAM2-K100R were transfected into HEK293T cells along with SIRT2-expressing plasmid, then the binding abilities between SIRT2 and PGAM2-WT or PGAM2-K100R were detected by Co-IP. G Exogenous Flag-PGAM2 and ubiquitin-expressing plasmids were transfected into HEK293T cells with or without SIRT2-expressing plasmid, then ubiquitination levels of PGAM2 was measured by Co-IP after treatment with 25 μM MG132 for 16 h. H PGAM2 was overexpressed in HEK293T and HCCLM3 cell lines along with SIRT2 or not, and cell lysates were used to examine the level of PGAM2 by western blot. I PGAM2 was measured by western blot in HCCLM3 cell line treated with 40 mM NAM or 20 μM SIRT2 inhibitor SirReal2 for 12 h. J, K Cell growth rates and colony formation abilities detection of HCCLM3 cell lines overexpressing Vector, PGAM2, SIRT2 alone or in combination. L Detection of cell apoptosis rates of HCCLM3 cell lines overexpressing Vector, PGAM2, SIRT2 alone or in combination. **p < 0.01, ***p < 0.001, ns no significance.