Figure 2.

PKM2deliversphosphorylated-Ser293 PDHA1 (p−PDHA1) to the nucleus. (A,B) HepG2 cells were stimulated with 100 nM insulin for 48 h (A) or 1, 24, and 48 h (B), p-PDHA1 and PKM2 changes were identified with immunohistochemistry. (C) HepG2 cells were treated with 100 nM insulin for 48 h, and cytosolic and nuclear fractions were separated, and then p-PDHA1 and PKM2 were determined by Western blotting. (D) si-PKM2 was transfected into HepG2 cells and insulin (100 nM for 48 h) was added to the cells, and p-PDHA1 levels were visualized with immunohistochemistry. Knockdown PKM2 levels were measured by Western blotting. (E) si-PDHA1 was transfected, then PKM2 was visualized with immunohistochemistry followed by insulin activation (100 nM for 48 h). PDHA1 silencing level was confirmed by immunoblotting. Statistic data were showed as * p < 0.05, ** p ˂ 0.01; *** p ˂ 0.001 and scale bar was 10 µm.