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. 2021 Sep 3;14:4347–4359. doi: 10.2147/JIR.S329091

Figure 4.

Figure 4

KAR mediates macrophage phenotype and function via aryl hydrocarbon receptor (AhR). (A) Mouse bone marrow-derived macrophages (BMDM) were generated as indicated and incubated in vitro with KAR (20 µg/mL) or PBS. AhR mRNA expression was examined using qRT-PCR. *P < 0.05, Student’s t test. (B–D) BMDM were generated from AhR+/+ or AhR−/- mice. Four groups of BMDM cultures were set up: LPS (100 ng/mL, Sigma-Aldrich, St. Louis, MO, USA)-activated AhR+/+ BMDM with PBS, LPS-activated AhR+/+ BMDM with KAR, LPS-activated AhR−/- BMDM with PBS and LPS-activated AhR−/- BMDM with KAR. After 2 days of culture, BMDM were harvested and qRT-PCR was performed to determine the expression levels of indicated genes (B and C). Four groups of BMDM were obtained as described in (B and C) and co-cultured with splenic naïve CD4+ T cells pre-labelled with carboxyfluorescein succinimidyl ester (CFSE) in the presence of anti-CD3/CD28. The ratio of T cells: BMDM was 4:1. After 5 days of co-culture, flow cytometry was performed to analyze (D) the dilution of CFSE intensity and (E) the expression of IFN-γ. (B–E) *P < 0.05, **P < 0.01, ***P < 0.001, one-way analysis of variance (ANOVA). Triplicates were carried out in each experiment. Representative results from one of three independent experiments were shown.