Figure 4.

FLV peptide rescued mitochondrial respiration deficits in M17 cells overexpressing D620N VPS35 mutant. (A) Mitochondria respiration activity was measured by Seahorse assay in M17 cells overexpressing vector (Ct) or VPS35 D620N mutant treated with scramble peptide or FLV peptide. Oxygen consumption (OCR) rate was measured before and after cells being sequentially exposed to oligomycin (inhibits ATP synthase, blocks oxygen consumption related to ATP synthesis), FCCP (uncoupler to assess maximal OCR), and antimycin A/rotenone (blocks electron flux through both complex I and II). (B–E) Quantification in these M17 cells of Basal OCR (B), Maximal OCR (C, OCR_FCCP), Respiratory control ratio (D, OCR_FCCP/OCR_Oligomycin), and Spare respiratory capacity (OCR_FCCP-OCR_Basal) were calculated after subtracting the non-mitochondrial respiration (E, OCR_{Antimycin A/rotenone}). Data are means ± s.e.m of 3 independent experiments. Statistics: one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test. *P < 0.05, **P < 0.01 and ***P < 0.001.