OC numbers and volumes in EBI2- and CH25H-deficient mice. (A) Two-photon microscopy of WT, EBI2-deficient, and CH25H-deficient femurs. Representative of at least five independent experiments. Bar, 50 µm. (B and C) OC total number (B) and total volume (C) in WT, EBI2-deficient, and CH25H-deficient femurs quantified from two-photon microscopy. Data are representative of more than five independent experiments. (D–F) Flow cytometric analysis of cells isolated from BM (D), peripheral blood (PB; E), and spleen (SP; F) of the indicated mice genotypes. MDP cells were gated as CD11b^loCSF1R⁺CD117⁺; cMoP cells were gated as Ly6C^hiCD11b^loCSF1R⁺CD117⁺; inflammatory monocytes (Inf.Mono.) were gated as CSF1R⁺Ly6C^hiCD11b⁺; patrolling monocytes (Pat.Mono.) were gated as CSF1R⁺Ly6C^int/loCD11b⁺; and neutrophils were gated as CSF1R⁻Ly6C⁺CD11b⁺. Data are representative of at least three independent experiments. Bars indicate the mean of WT (white, n = 14), Ebi2^−/− (green, n = 12), and Ch25h^−/− (blue, n = 12) mice; circles depict individual mice. *, P < 0.05; **, P < 0.01 by unpaired Student’s t test.