Figure 4.

Suppression of LPS-induced NO metabolites and PGE₂ production by erastin in bone marrow-derived macrophages (BMDMs). (A) iNOS and (B) COX2 mRNA levels were analyzed by real-time qPCR in BMDMs after stimulation with 1 μg/mL of LPS for 24 h with or without 5, 10, or 20 μM of erastin. (C) Protein expression of inflammatory enzymes (iNOS and COX2) in LPS-induced BMDMs with or without the indicated dose of erastin (1, 5, 10, or 20 μM) were examined by western blotting. Cell lysates were harvested at 24 h after LPS treatment with or without indicated dose of erastin; β-actin was used as a loading control. (D) NO metabolite secretion was determined using Griess reagent and (E) PGE₂ secretion was analyzed by Prostaglandin E₂ ELISA Kit in BMDM culture medium. BMDMs were induced with 1 μg/mL of LPS for 24 h with or without erastin at various doses (5, 10, 20 μM). Graphs represent the mean of three independent experiments. Statistical analyses were performed using paired two-tailed Student’s t-test. * p < 0.05, # p < 0.01.