Fig. 4
Mutation of key lysine residues blocks Drp1 SUMOylation and rescues mitochondria-mediated NPC apoptosis. a MAPL overexpression-mediated SUMOylation of Drp1 was analysed via co-IP in NPCs transfected with WT-Drp1 or the 4KR-N, 4KR-C and 8KR mutants. b Western blots showing the levels of total Drp1, MAPL, cleaved caspase-3, Bax, and Bcl-2 in NPCs transfected with the WT-Drp1 or 8KR plasmid after MAPL was overexpressed. c Western blot analysis of Drp1 and MAPL levels in the cytosolic fraction and mitochondrial compartment of NPCs after MAPL overexpression and lysine mutation of Drp1. d The translocation of Drp1 in NPCs was observed using MitoTracker Red staining and immunofluorescence staining for Drp1. e Changes in the morphology of mitochondria in MAPL-overexpressing NPCs transfected with WT-Drp1 or 8KR, as shown by MitoTracker Green staining. f JC-1 staining and flow cytometry were used to detect the ΔΨm in NPCs transfected with WT-Drp1 or 8KR after MAPL overexpression. g Flow cytometry with Annexin V-FITC/PI staining was used to detect the apoptotic rate of NPCs. (n = 3; *P < 0.05, **P < 0.01, and ***P < 0.001)
