Fig. 5: MCL-1 is required for mitophagy induced by oxygen-glucose deprivation.

a HEK293T-mt-Keima cells were infected with lentiviral particles encoding MCL-1 shRNA for 24 h and treated with oxygen-glucose deprivation (OGD) for 5 h. Mitophagy levels were analyzed by one-way ANOVA (data represent mean ± S.E.M.; n = 4. ****p < 0.0001, ns, not significant). shNC: scrambled shRNA. b HEK293T-mt-Keima cells were infected with lentiviral particles encoding MCL-1 shRNA for 24 h and treated with OGD for 5 h. Cell lysates were immunoblotted with indicated antibodies. The numbers under the blots represent the gray scale quantification (Cox II/Tubulin, Tim23/Tubulin). shNC: scrambled shRNA. c HEK293T cells were infected with lentiviral particles encoding MCL-1-shRNA or control-shRNA for 24 h, treated with OGD for 5 h. Cells were fixed and stained with indicated antibodies. Green channel indicated cells were infected successfully. Scale bar, 5 μm; insets: scale bar, 2 μm. shNC: scrambled shRNA. d HEK293T-MCL-1-knockdown cells were transfected with MCL-1 wild-type or indicated mutants for 24 h, treated with OGD for 5 h. Cells were fixed and stained with indicated antibodies, following by fluorescence microscopy analysis. Scale bar, 5 μm; insets: Scale bar, 2 μm. e HEK293T-MCL-1-knockdown cells were transfected with MCL-1 WT or indicated mutants for 24 h, treated with OGD for 5 h. Cell lysates were immunoblotted with indicated antibodies. The numbers under the blots represent the gray scale quantification (Tom20/Tubulin, Tim23/Tubulin). f HEK293T cells were co-transfected with MCL-1-3xFlag and HA-LC3A for 24 h, treated with OGD for 5 h, and the interaction of Flag-tagged MCL-1 with HA-LC3A and endogenous Bax was analyzed by immunoprecipitation. Source data are provided as a Source Data file.