Extended Data Fig. 5: PKCβII did not affect the protein level of GPX4 or cellular divalent iron levels.
From: PKCβII phosphorylates ACSL4 to amplify lipid peroxidation to induce ferroptosis
(a) An immunoblot showing the expression of GPX4 in MDA-MB-231 cells subjected to the indicated treatments for 18 h. (b) Cellular divalent iron levels were assayed in the indicated MDA-MB-231 cells. (c) Cellular divalent iron levels were assayed in MDA-MB-231 cells subjected to the indicated treatments. Go6983, 5 μM; Enza, 5 μM. (d) The potential phosphorylation sites of ACSL4 predicted by GPS and the SCANSITE program. (e) An immunoblot showing the levels of ACSL4 in the indicated cells. (f-i) Endogenous ACSL4 was immunoprecipitated from cells subjected to the indicated treatments for 16 h, followed by immunoblotting using an ACSL4 phospho-T328 antibody. For MDA-MB-231 cells, erastin, 5 μM; RSL3, 2.5 μM. For HT1080 cells, erastin, 4 μM; RSL3, 50 nM. b, c, the mean±s.d. of n = 3 independent experiments are shown. Statistical analysis was performed using an unpaired two-tailed t test. Data in a, e-i, are representative of n = 3 biologically independent experiments.
