Fig. S6.

OPTN single- and double-Ser mutations have limited effect on translocation and mitophagy. (A) Western blot of WT or PentaKO HeLa cells with or without stable expression of GFP–OPTN for OPTN expression levels. (B) Western blot of mCherry–Parkin-expressing pentaKO cells reconstituted with GFP–OPTN WT and indicated mutants as used in C and D and Fig. 5A. Western blot shows equal expression levels. (C and D) Representative images of mCherry–Parkin pentaKO cells expressing OPTN WT or OPTN mutants as indicated were untreated or treated with antimycin A or oligomycin (AO) for 0.5 h (C) and 3 h (D) and immunostained for TOM20. (E) Cells from B–D were treated with AO for the indicated times and lysates immunoblotted for pTBK1.