Figure EV3. Oxidation of NDP52 facilitates PINK1/Parkin mitophagy with no role in mitophagy triggered by iron chelation.

• A
  Fluorescence images of YFP‐Parkin in HeLa WT, PentaKO, PentaKO + NDP52 WT and PentaKO + NDP52 Mut cells stably expressing YFP‐Parkin and mt‐mKeima, in the same fields and conditions as Fig 4A.
• B
  Fluorescence microscopy images and quantification of mitophagy of HeLa WT, PentaKO, PentaKO + NDP52 WT and PentaKO + NDP52 Mut cells stably expressing YFP‐Parkin and mt‐mKeima were treated with 1 mM DFP for 24 h.
• C
  Fluorescence microscopy images and quantification of mitophagy of HeLa WT, PentaKO, PentaKO + NDP52 WT and PentaKO + NDP52 Mut cells stably expressing YFP‐Parkin and mt‐mKeima, pre‐treated with or without 500 nM MitoQ for 16 h and treated with 10 μM G‐TPP for 8 h.
• D–F
  HeLa PentaKO + NDP52 WT cells were pre‐treated with or without MitoQ for 16 h and treated with G‐TPP for 5 h or AO for 2 h, followed by immunoblotting for NDP52 in non‐reducing conditions (D), MitoSOX staining (E) and mitochondrial membrane potential assay by using TMRM and Mitotracker Green (MTG) staining (F).

Data information: Data are mean ± s.e.m. (B, C, E, F) or displayed as cell popular violin plots (C). P values were calculated by one‐way ANOVA followed by Sidak test on three independent experiments (B, C, E, F). *, P < 0.05; **, P < 0.01; ***, P < 0.001; §§, P < 0.01; §§§, P < 0.001 (relative to MitoQ‐untreated condition); ns (non‐significant). Scale bars: 20 μm (A, B, C, E, F).

Source data are available online for this figure.