Extended Data Figure 3: TBK1 activates OPTN in PINK1/parkin mitophagy.
From: The ubiquitin kinase PINK1 recruits autophagy receptors to induce mitophagy
a, Representative images of untreated mCh–parkin cells and merged images of treated cells as indicated immunostained for DNA. See Fig. 2a for anti-DNA and DAPI images of treated samples (n = 3). b, Cell lysates from wild-type, NDP52/OPTN DKO, OPTN knockout and NDP52 knockout cells with or without mCh–parkin expression were immunoblotted for TBK1 activation. c, Cell lysates from wild-type and PINK1 knockout cells without parkin expression were immunoblotted for TBK1 activation (S172 phosphorylation). d, Confirmation of TBK1/NDP52 DKO, TBK1/OPTN DKO and TBK1 knockout by immunoblotting. e, Knockout cell lines from d were immunostained for TOM20 (n = 3). f, Representative images of untreated mCh–parkin wild-type and knockout cells, and merged images of treated cells as indicated were immunostained for DNA. See Fig. 2g for anti-DNA/DAPI images treated samples (n = 3). g, TBK1/NDP52 DKO cells rescued with GFP–TBK1 wild-type or K38M, or GFP–OPTN(S177D) and were assessed by immunoblotting. h, Cells in g were assessed by immunoblotting. i, Quantification of COXII levels in h displayed as mean ± s.d. from three independent experiments. **P < 0.005 (one-way ANOVA). Scale bars, 10 μm.
