Extended Data Fig. 8. ERK2 overexpression in RPE-1 cells increases phosphorylated ERK2, NOX activity, and the speed of ferroptotic trigger waves.
a, Genetic modulation of NOX signalling by ERK2 overexpression. b, Western blot analysis of overexpressed ERK2 (ERK2-P2A) and its phosphorylated form (phospho-ERK2-P2A) with or without erastin treatment (10 μM) in ERK2-overexpressing and control RPE-1 cells. The size of ERK2-P2A is bigger than wild type ERK2 and co-migrates with ERK1 due to its fusion to P2A peptide. For blot source data, see Supplementary Fig. 1. c, Relative NOX activity measured in ERK2-overexpressing and control RPE-1 cells. Data represent mean ± s.d. with four technical repeats. NOX activity of ERK2-overexpressing cells is significantly different from that of control cells (* two-sided Wilcoxon rank-sum test P = 0.0286). d, e, ERK2 overexpression increases the speed of ferroptotic trigger waves. Kymographs representing ferroptosis propagation in control cells (d) and ERK-overexpressing cells (e). Data shown (b-e) are representative of three biological repeats.