Figure 6.

The Drp1‐dependent fission and mitophagy facilitated HK2 separation from the mitochondria and the inhibition of glycolysis. SMMC‐7721 cells were treated with indicated concentrations of CTB (0, 1, 2 and 4 μmol/L), Mdivi‐1 (50 μmol/L) or rapamycin (100 nmol/L) for 24 h. (A) HK activity was detected by Hexokinase Activity Detection Kit. (B‐C) Mitochondrial and cytosolic fractions were isolated. Western blot analysis showed the protein expression of HK2. (D) Representative fluorescence microscope images of SMMC‐7721 cells labelled with DAPI, HK2 antibody and Mito‐Tracker Green. Scale bar: 50 μm. (E) Glucose consumption was detected by glucose assay kit. (F) Production of lactic acid was assayed by Lactic Acid Production Detection kit. (G) ATP content was detected by the ATP Assess Kit. Data were presented as mean ± SD (n = 5); significance: *P < .05, **P < .01 and ***P < .001 vs control; ^# P < .05, ^## P < .01 and ^### P < .001vs CTB treatment