Fig. 3.

DAT does not affect autophagic flux and sensitizes cells to ferroptosis through a mechanism independent of autophagy. a Ferroptosis was induced by erastin with or without the addition of DAT and BafA1. Autophagic activity was assessed by western blot analysis of LC3-I/LC3-II and p62 protein expression. b Immunofluorescence analysis of GFP-LC3-II accumulation after erastin-induced ferroptosis with or without BafA1. The presence of puncta indicates LC3-II. For a–b, MEFs were treated with erastin for 8 h and HT1080 for 10 h. c Percentage cell death was quantified by flow cytometric analysis of PI-positive cells in Atg5-KO MEFs, or isogenic Atg5-KO cells reconstituted with ectopic Atg5 expression. For a–c, cells were treated with 10 μM DAT, 1 μM erastin and 20 nM BafA1