Fig. 3.

Ginsenoside CK inhibits mitochondrial dynamics imbalance and damage by inhibiting the ubiquitination of Mfn2 by Mul1. (A) The expression of Mul1 and mitochondrial dynamics-related proteins in cytoplasm or mitochondria was detected by western blot; β-Actin and TOM20 were the loading controls for cytosolic and mitochondrial proteins, respectively. (B) After Mul1 or Mfn2 was immunoprecipitated, the binding and ubiquitination level of Mfn2 in mitochondrial proteins were detected by western blot analysis; 10% of the lysed mitochondrial proteins in the co-IP experiment were used as input control. (C) The expression of Mul1, Mfn2, and DRP1 were detected by western blot and quantitatively analyzed by ImageJ software. (D) The level of mitochondrial ROS (MitoSox) was determined by confocal microscope; scale bar = 20 μm. Hochest 33254 was used as a nuclear counterstain.