Extended Data Fig. 1: Energy stress inhibits ferroptosis.
From: Energy-stress-mediated AMPK activation inhibits ferroptosis
a, Immunoblot analysis in MEFs treated with 2 μM erastin (16 h), cystine free media (8 h), 100 nM RSL3 (16 h), or 1 μM of staurosporine (STS, 2 h). b, Cell death measurement in MEFs treated with 2 μM erastin and cell death inhibitors for 16 h. Ferr-1: 1 μM ferrostatin-1; DFO: 100 μM deferoxamine; Z-V: 20 μM Z-VAD-FMK; Nec: 2 μM necrostatin-1; NAC: 5 mM N-acetyl cysteine. c, Cell death measurement in MEFs cultured in 25 or 0 mM glucose-containing medium with erastin and/or Ferrostatin-1 for the indicated times. d, Immunoblot analysis in MEFs treated as in a, or glucose starvation for 48 h. e, Cell death measurement in MEFs cultured in cystine free media with cell death inhibitors for 8 h. f, g, Intracellular ATP levels (f) and cell death measurement (g) in MEFs cultured with the indicated concentrations of glucose for 16 h. h–k, The measurement of cell death (h, j) and lipid peroxidation (i, k) in Caki-1 or BJ cells. Cells were treated with A769662 (200 μM), AICAR (2 mM), 2DG (5 mM), 0 mM glucose with simultaneous treatment of 2 μM erastin for 24 h (cell death) and 16 h (Lipid peroxidation). P values correspond to the comparison between control and each treatment in red bars. l, Immunoblot showing the levels of AMPK T172 phosphorylation. MEFs cells were treated as in h–k and compound C (5 μM) for 16 h. m, Cell death measurement in AMPK WT and DKO MEFs treated with 2 μM erastin and 2 mM of AICAR for 16 h. n, Cell death measurement in AMPK WT and DKO MEFs treated with erastin at the indicated concentrations for 16 h. P values correspond to the comparison between AMPK WT and DKO at indicated erastin concentrations. Data show the mean ± s.d., n = 3 independent experiments. Statistical analysis was performed using unpaired, two-tailed t-test. Numerical source data are provided in Source Data Extended Data Fig. 1. Scanned images of unprocessed blots are shown in Source Data Extended Data Fig. 1.
