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. 2016 Aug 12;26(9):1021–1032. doi: 10.1038/cr.2016.95

Figure 2.

Figure 2

Ferroptosis inducer activates autophagy. (A, B) Erastin induced LC3 conversion in MEFs (A) and HT1080 cells (B). Cells seeded in six-well dishes were treated with the 0.5 μM (A) or 5 μM (B) erastin for the indicated times. 20 nM BafA1 was added 2 h before cell harvest. Cell extracts were analyzed by western blot using antibodies against the indicated proteins. The accumulation of LC3II (faster migrating form) relative to LC3I (slower migrating form) is indicative of the induction of autophagy. (C, D) Erastin induced GFP-LC3 puncta formation in MEFs (C) and HT1080 cells (D). Cells stably expressing GFP-LC3 grown on glass cover slips were either left untreated or treated with 0.5 μM (C) or 5 μM (D) erastin for 7 h. Cells were then fixed with 3.7% PFA, processed for imaging, and visualized under the confocal microscope using the 60× magnification objective.