Fig. 5: APOH inhibits ferroptosis by activating the AKT-SREBP1 pathway.

A APOH induced the phosphorylation of AKT (p-AKT) in MEFs. Western blot analysis was performed to measure the p-AKT, AKT, and APOH. MEFs cultured in Scm were serum-starved for 12 h and then treated with the indicated concentration of APOH, or 5% or 10% of Rcm or Scm for 30 min. B PI3K/AKT inhibitor MK-2206 prevented the p-AKT induced by APOH. MEFs cultured in Scm were serum-starved for 12 h and then treated with 10 μg/mL APOH or heat-inactivated APOH for the indicated time with or without MK-2206 (5 μM). Western blot analysis was performed to measure p-AKT and AKT. C Cell viability of MEFs cultured in Scm changed medium to Rcm or not, treated with or without LY294002 (LY, 12 μM) for 12 h before treatment with different doses of RSL3 and/or Lip-1 (1 μM) for 24 h. D Cell viability of MEFs cultured in Scm and then changed medium to 5% Scm + 5% Rcm, 5% Scm + APOH (120 μg/mL) or not, treated with or without LY (12 μM) for 12 h before treatment with different doses of RSL3 and/or Lip-1 (1 μM) for 24 h. E Cell viability of MEFs cultured in Scm changed medium to 5% Scm + 5% Rcm, 5% Scm + WT-APOH (120 μg/mL), 5% Scm + DDV-APOH (120 μg/mL) or not, treated with different doses of RSL3 for 24 h. F Cell viability of MEFs cultured in Scm changed medium to Scm + ApoHinfer (Scm adding 300 μg/mL ApoHinfer) or not for 24 h, before treated with or without 0.4 μM RSL3 or 0.2 μM ERS2 for 24 h. Western blot is representative of three biological replicates (A, B). Data are presented as mean ± s.d., n = 3 independent repeats. Unpaired, two-tailed t-test; **P < 0.01, ****P < 0.0001. NS not significant.