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. 2019 Dec 14;8(12):2210. doi: 10.3390/jcm8122210

Figure 5.

Figure 5

Erastin suppresses the induction of inflammatory cytokines in LPS-induced bone marrow-derived macrophages (BMDMs). Transcription levels of (A) TNF-α and (B) IL-1β after activation of BMDMs with 1 μg/mL of LPS for 24 h with or without 5, 10, or 20 μM of erastin were measured using real-time qPCR. (C) Protein expression of inflammatory cytokines (TNF-α and IL-1β) in LPS-induced BMDMs with or without the indicated dose of erastin (1, 5, 10, or 20 μM) were analyzed by western blot. 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) TNF-α, and (E) IL-1β secretion levels after treating BMDMs with 1 μg/mL of LPS for 24 h with or without erastin at various doses (5, 10, 20 μM) in culture medium were determined by ELISA assay. Time-dependent secretion of (F) TNF-α and (G) IL-1β from BMDMs following stimulation with 1 μg/mL of LPS for 8, 16, and 24 h with or without 20 μM of erastin was assayed using ELISA. Graphs represent mean of three independent experiments. Statistical analyses were performed using paired two-tailed Student’s t-test. * p < 0.05, # p < 0.01.