Extended Data Fig. 4: PKCβII activation is essential for ferroptosis.
From: PKCβII phosphorylates ACSL4 to amplify lipid peroxidation to induce ferroptosis
(a-b) Cell death measurements in MDA-MB-231 (a) and HT1080 (b) cells subjected to the indicated treatments. (c) An immunoblot showing the expression of membrane-PKCα, βI, γ, δ, ζ in MDA-MB-231 cells treated with erastin at the indicated concentration for 16 h. (d) Lipid peroxidation measurement in the indicated MDA-MB-231 cells or HT1080 cells treated with erastin or RLS3 for 20 h. For MDA-MB-231 cells, erastin, 5 μM; RSL3, 2.5 μM. For HT1080 cells, erastin, 4 μM; RSL3, 50 nM. The mean ± s.d. of n = 3 independent experiments are shown. Statistical analysis was performed using an unpaired two-tailed t test. (e,f) An immunoblot showing the expression of membrane-PKCβII in MDA-MB-231 (e) or HT1080 cells (f) subjected to the indicated treatments for 16 h. For MDA-MB-231 cells, erastin, 5 μM; RSL3, 2.5 μM. For HT1080 cells, erastin, 4 μM; RSL3, 50 nM. (g,h) Endogenous PKCβII was immunoprecipitated from MDA-MB-231 (g) and HT1080 (h) cells subjected to the indicated treatments for 16 h, followed by immunoblotting using a phospho-Ser/Thr antibody. For MDA-MB-231 cells, erastin, 5 μM; RSL3, 2.5 μM. For HT1080 cells, erastin, 4 μM; RSL3, 50 nM. a, b, d, the mean ± s.d. of n = 3 independent experiments are shown. Statistical analysis was performed using an unpaired two-tailed t test. Data in c, e-h, are representative of n = 3 biologically independent experiments.
