5.

Hexokinase (HK) II increases oxygen consumption rate (OCR) of cells and promotes cell proliferation by phosphorylating S293 of the alpha subunit of pyruvate dehydrogenase (PDHA1). (A) OCRs were detected (bottom) for ctrl and HKII KD cells; (B) OCRs were detected (bottom) for ctrl and HKII overexpressed cells (with or without 3BP, 5 μmol/L). OCR under oligomycin, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and antimycin A/rotenone (R/A) treatments, respectively; (C) Growth curves of control HEK293T cells, HKII overexpressing HEK293T cells and HKII knockdown HEK293T cells were determined; (D) Growth curves of control HEK293T cells, HKII overexpressing HEK293T cells, HKII & PHDA1 overexpressing HEK293T cells and HKII & S293A overexpressing HEK293T cells were determined; (E) PDHA1 was knocked down in HEK293T cells and then PDHA1 and S293A and HKII were re-introduced into the cells. Growth curves of siPDHA1 HEK293T cells, PDHA1 overexpressing siPDHA1 HEK293T cells, HKII & PHDA1 overexpressing siPDHA1 HEK293T cells and HKII & S293A overexpressing siPDHA1 HEK293T cells were determined; (F) PDHA1 was knocked down in HEK293T cells and then PDHA1 was transfected into cells and finally these cells were transfected with HKII or HKII mutation and growth curves were determined. *, P<0.05; **, P<0.01; ***, P<0.001.