Parkin-mediated mitophagy in HeLa cells

All cells were maintained at 37°C and 5% CO₂ under humidified conditions. HeLa cells were obtained from ATCC. Control primary fibroblasts were from Cell Applications, Inc. PINK1 Q456X fibroblasts have been described before [31 (link), 45 ]. HeLa cells and fibroblasts were cultured in Dulbecco’s modified eagle medium (DMEM, Invitrogen) containing 10% FBS (BioWest). Fibroblast medium was supplemented with 1% non-essential amino acid and 1% Penicillin/Streptomycin (both Invitrogen). HeLa cells stably expressing EGFP-Parkin, untagged Parkin, 3xFLAG-Parkin C431S or the EGFP-Parkin and mitoKeima have been described before [16 (link), 21 (link), 25 (link)]. iNeurons were generated as described [16 (link)].