Extended Data Figure 2: OPTN, NDP52 and TAX1BP1 triple knockout analysis and disease-associated mutations.

a, b, Knockout cell lines with or without mCh–parkin expression were immunoblotted (a) and COXII levels were quantified (b). c, A panel of human tissue lysates was immunoblotted. d, Expression of wild-type or mutant GFP–OPTN in mCh–parkin pentaKO cells. e, Quantification of cells in f. More than 100 cells per condition. f, Representative images of mCh–parkin pentaKO cells expressing GFP–OPTN mutants immunostained for TOM20 (n = 3). g, PentaKOs expressing mCh–parkin were rescued with wild-type or mutant GFP–OPTN, analysed by immunoblotting. See Fig. 2e for quantification of COXII. h, Expression of wild-type or mutant GFP–NDP52 in mCh–parkin pentaKO cells. i, Quantification of cells in j. More than 100 cells per condition. j, Representative images of mCh–parkin pentaKOs expressing wild-type or mutant GFP–NDP52 were immunostained for TOM20 (n = 3). k, PentaKO cells expressing mCh–parkin rescued with wild-type or mutant GFP–NDP52 were analysed by immunoblotting. See Fig. 2f for quantification of COXII. Data are mean ± s.d. from three (b and i) and two (e) independent experiments. ***P < 0.001 (one-way ANOVA). Scale bars, 10 μm.