MDVs are not essential for misfolded protein clearance from mitochondria. (A) Tet ON: ΔOTC-expressing HeLa cells also expressing YFP-Parkin with or without a Drp1 KO background were treated with 20 nM control, syntaxin 17, or pink1 RNAi for 24 h and then treated with DOX for 48 h or 48 h with a 24-h washout of DOX, and cells were processed for Western blot. (B) Quantification of Western blots as described in A expressed as the percentage of ΔOTC levels relative to ΔOTC levels after 48 h DOX treatment normalized to Hsp90 levels. n ≥ 3. (C) HeLa cells expressing YFP-Parkin with or without a Drp1 KO background and with or without a syntaxin 17 KO background were processed for Western blot analysis. The asterisk indicates a nonspecific band. (D) HeLa cells expressing YFP-Parkin with a Drp1 KO background and with or without a syntaxin 17 KO background were treated with actinonin for 16 h with and without 400 nM bafilomycin treatment and then were processed for Western blot analysis. (E) Quantification of Western blots as described in D expressed as the percentage of control TIM23 or Hsp60 levels normalized to Hsp90 levels. n ≥ 3. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Error bars indicate SD.