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. 2022 Aug 9;13:938944. doi: 10.3389/fimmu.2022.938944

Figure 1.

Figure 1

Blockade of endogenous EPO impaired re-programming of endotoxin-tolerant macrophages. (A, B): In vitro cultured BMDMs from WT C57BL/6 mice (A) or mouse RAW264.7 macrophages (B) were incubated with LPS (100 ng/ml) for 0, 6, 12, and 24 h, and mRNA levels of Hif1a, Epo, and Epor in macrophages were measured by qRT-PCR (n = 3). (C, D): In vitro cultured RAW264.7 macrophages were pretreated with PBS (non-tolerant), LPS (100 ng/ml) (tolerant), or LPS (100 ng/ml) + anti-EPO-16 (10 or 20 μg/ml) for 24 h. Then, cells were washed with PBS twice. Subsequently, cells were given with a secondary LPS stimulation (10 ng/ml) for 6 h, and the mRNA levels of Il1b, Il6, Tnfa (C), Cnlp, Marco, and Vegfc (D) were measured by qRT-PCR (n = 3). Data are representative of three independent experiments. Results were expressed as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (one-way ANOVA with Tukey’s post-hoc test for multiple comparisons).