Fig. 1: Crizotinib suppresses ferroptosis in various cell lines independently of ALK expression.

A (R)-CRZ protects HT-1080 cells from RSL3-induced ferroptosis. HT-1080 cells were treated with RSL3 (0.25 μM) with or without (R)-CRZ (1 μM) added. Cell morphology was recorded at 24 h after drug treatment. B (R)-CRZ (1 μM) inhibits RSL3 (0.25 μM)-induced ferroptosis in both SK-HEP-1 and HT-1080 cell lines. Fer-1 (1 μM) was used as a positive control. Cell viability was measured after 10 h of treatment. C (R)-CRZ inhibits ferroptosis induced by RSL3 or ML210. (R)-CRZ (1 μM) and Fer-1 (1 μM) were used in the experiments. D (R)-CRZ (1 μM) inhibits ferroptosis induced by PACMA31 (1 μM) or RSL3 (1 μM) in multiple cell lines. Cell viability was assayed after 10 h of treatment. E Other ALK inhibitors do not inhibit ferroptosis. (R)-CRZ (3 μM), lorlatinib (3 μM), alectinib (3 μM), Fer-1 (1 μM) and ceritinib (1 μM) were used in the experiments. Cell viability was measured after 8 h of treatment. F Cell lines rescued from ferroptosis by (R)-CRZ did not show ALK expression as validated by Western blot. SU-DHL-1 and Karpas-299 cell lines were used as positive controls for ALK expression. G Some other RTK inhibitors do not affect ferroptosis. All tested compounds were used at a concentration of 3 μM in the experiment. H Knocking-down of AXL in 786-O cells do not affect cell sensitivity to ferroptosis. Cell viability was measured after 17 h of treatment. I (R)-CRZ (1 μM) do not affect GPX4 and SLC7A11 protein expression assayed by Western blot. J (R)-CRZ (3 μM) inhibits spontaneous ferroptosis in GPX4 KO monoclonal HT-1080 cell lines. Cell viability was measured after 12 h of treatments. Data in all quantitative statistical charts of Fig. 1 are presented as the mean ± S.D., n = 3 independent repeats.