Cold stress‐induced ferroptosis involves the ASK1‐p38 pathway

A

Immunoblot of endogenous phospho‐ASK and phospho‐p38 signals in HT‐1080 cells after 4 h of treatment with 1 μM RSL3 in the presence or absence of 1 μM ferrostatin‐1. RSL3 administration was performed immediately after ferrostatin‐1 treatment.

B, C, E, F

HEK293A (B, C) or HT‐1080 (E, F) cells were transfected with siRNAs followed by erastin (B, E) or RSL3 (C, F) treatment. Cells were reseeded a day prior to erastin or RSL3 treatment in order to match the number of cells among each sample. Cells were treated with erastin or RSL3 at the indicated concentration for 24 h. Cell viability was measured using the Cell Counting Kit‐8. Mean ± SEM, N = 5 (B, F), 3 (C, E). Two‐way ANOVA followed by Tukey's multiple comparisons test. See also Appendix Table S2H–K.

D, G

Immunoblots of endogenous ASK1 and α‐tubulin in HEK293A (D) or HT‐1080 (G) cells for confirming the knockdown efficiency of the samples used in (B and C), and (E and F), respectively.

Figure EV4. ASK1 does not affect ferroptotic cell death in HEK293A and HT‐1080 cells.