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. 2024 Mar 29;10(13):eadk1200. doi: 10.1126/sciadv.adk1200

Fig. 2. AS is a common inhibitor of ferroptosis.

Fig. 2.

(A and B) Cell viability analysis of MDA-MB-231 cells (A) or HT22 cells (B) pretreated with or without AS for 1 hour and then stimulated with different concentrations of RSL3 for 8 hours. (C and E) Lipid peroxidation was analyzed by flow cytometry in MDA-MB-231 cells (C) or HT22 cells (E) pretreated with AS or Fer-1 for 1 hour and then stimulated with 1 μM RSL3 for 8 hours in the presence of C11-BODIPY. (D and F) The percentage of C11-BODIPY–positive MDA-MB-231 cells (D) or HT22 cells (F) pretreated with AS or Fer-1 for 1 hour and then stimulated with 1 μM RSL3 for 8 hours in the presence of C11-BODIPY. (G and H) Cell viability (G) or LDH release (H) assay of HT-1080 cells pretreated with or without AS for 1 h and then stimulated with 1 μM RSL3, 20 μM erastin, or 40 μM FIN56. (I and J) Cell viability (I) or LDH release (J) assay of MDA-MB-231 cells pretreated with or without AS for 1 hour and then stimulated with 1 μM RSL3, 20 μM erastin, or 40 μM FIN56. (K and L) Cell viability (K) or LDH release (L) assay of MDA-MB-231 cells pretreated with or without AS for 1 hour and then stimulated with 1 μM RSL3, 20 μM erastin, or 40 μM FIN56. (M) CRISPR-Cas9–generated GPX4-knockout HT-1080 cells were treated with or without AS or Fer-1. The expression of GPX4 was detected by immunoblotting (left), and the cell viability was detected by CCK-8 assay (right). Data are the mean ± SEM, n = 3 biologically independent experiments [(D) and (F) to (M)]. Statistical analysis was performed using an unpaired two-tailed Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.