Figure 3.

Mitophagy regulators modulate extracellular mitochondria release. (A) Representative western blot of TOMM20, HSPD1, OPA1, CI-NDUFB8, CII-SDHB, CIV-MT-CO1 and CV-ATP5F1A relative to ACTB in CM-derived pellet collected following extended non-treated condition (24 h). (B) Representative western blot of TOMM20 in CM-derived pellet collected following brief DMSO and rotenone-treated condition (4 h). PRKN overexpression suppresses extracellular mitochondria release under extended non-treated condition (24 h) and brief DMSO and rotenone-treated condition (4 h). (C) PRKN knockdown elevates extracellular mitochondria release under extended non-treated condition (24 h) and (D) brief DMSO and rotenone-treated condition (4 h). (E) Quantification of total number of healthy and depolarized mitochondria released by DMSO and rotenone-treated control and PRKN knockdown cells. One-way ANOVA, Tukey’s test, n = 3 for each group, * p < 0.05. (F) Representative western blot and densitometry showing TOMM20 relative to ACTB in CM-derived pellet of control, BNIP3L/NIX and BNIP3 overexpressing PC12 cells following extended non-treated condition. (G) Representative western blot and densitometry showing TOMM20 relative to ACTB in CM-derived pellet of HeLa WT, ATG3 KO, ATG5 KO, ATG7 KO, ATG9 KO, ATG13 KO, ATG14 KO, RB1CC1/FIP200 KO cells following extended non-treated condition. One-way ANOVA, Dunnett’s test, n ≥ 3 for each group, * p < 0.05