GPR183 is dispensable for macrophage differentiation in vitro and for acquisition of homeostatic alveolar macrophage metabolism. (A) Metabolism of alveolar macrophages isolated from competitive Gpr183^+/+/Gpr183^−/− bone marrow chimeras 9–11 wk after bone marrow transfer. Metabolic dependencies and capacities were determined by SCENITH (Arguello et al., 2020) as described in the Materials and methods. AAO, amino acid oxidation; 2-DG, 2-deoxy-D-glucose; FAO, fatty acid oxidation; Oligo, oligomycin; Puro, puromycin. Data are represented as mean ± SEM. Data are combined from two independent experiments. For each experiment, BAL cells from two to three chimeric mice were pooled. In one of the two independent experiments, a sufficient number of cells allowed for two technical replicates. (B and C)In vitro differentiation of Gpr183^+/+ and Gpr183^−/− macrophages. Bone marrow cells from Gpr183^+/+ and Gpr183^−/− mice were mixed 1:1 and cultured with the indicated cytokines to generate macrophages in the absence or presence of 7α,25-dihydroxycholesterol (7α,25-OHC). The ratio of Gpr183^+/+/Gpr183^−/− macrophages was normalized to the bone marrow input of each independent experiment. Data are represented as mean ± SEM. Data are pooled from three independent experiments with a total of n = 3 mice per genotype and two technical replicates per experiment. Panel B was adapted from Servier Medical Art.