Skip to main content
. 2022 Mar 24;17(3):e0265738. doi: 10.1371/journal.pone.0265738

Fig 2. Hypoxia induced macrophage migration inhibitory factor (MIF) mRNA expression and protein production via the hypoxia inducible factor (HIF) pathway.

Fig 2

A. The effects of ML228, a HIF activator, on MIF mRNA expression in primary cultured astrocytes (PCAs). PCAs were incubated for 48 h in 0, 0.25, 0.5, or 1.0 μM ML228. MIF mRNA expression was analyzed by qRT-PCR. The values are shown as the ratio of MIF mRNA to beta-actin (ACTB) mRNA (Dunnett’s test vs. vehicle; n = 3). B. The time-dependent effects of hypoxia on MIF mRNA expression in PCAs. PCAs were incubated for 3, 6, 12, 24, or 48 h in 21.0% or 0.1% O2. MIF mRNA expression was analyzed by qRT-PCR. The values are shown as the ratio of MIF mRNA to ACTB mRNA (Student’s t-test; n = 5–6). C. The effects of the HIF inhibitor YC-1 on hypoxia-induced MIF mRNA expression in PCAs. PCAs were incubated for 6 h in 21.0% or 0.1% O2 and treated with 0 or 10 μM of YC-1. MIF mRNA expression was analyzed by qRT-PCR. The values are shown as the ratio of MIF mRNA to ACTB mRNA (Tukey’s test; n = 6). D. The effects of hypoxia on MIF protein production in cell lysate of PCAs. PCAs were incubated for 24 or 48 h in 21.0% or 0.1% O2. The MIF protein level per 1 μg of total protein was analyzed using a MIF enzyme-linked immunosorbent assay (ELISA) (Student’s t-test; n = 6). The data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01, or ***p < 0.001.