Extended flow cytometry characterization of Mφs and monocytes in the BM of naive and 5 week posttransplant MacGreen mice. Representative flow cytometry analysis of BM myeloid populations, extending data provided in Figure 1. (A) Naive (no-Tx) MacGreen BM stained with anti-F4/80 and anti-Ly6G antibodies (right plot) demonstrating specificity of staining and gating of F4/80⁺Ly6G^neg monocyte-Mφ–enriched cells. (B) Naive C57BL/6 BM stained with appropriate isotype control (left plot) or anti-VCAM-1 antibody (right plot) demonstrating specificity of staining and lower limit of detection for GFP transgene expression. (C-I) Representative flow cytometry analysis (n = 3-13) of BM from no transplant and 5 weeks posttransplant MacGreen mice demonstrating isotype control staining in 5 weeks posttransplant BM (C), VCAM-1 and GFP staining in no-Tx and 5 weeks posttransplant (D), and extended ER-HR3, Ly6C, MerTK, and TIM-4 expression in no-transplant Mφs (E), no-transplant monocyte (F) , donor Mφ (G), donor monocyte (H), and recipient Mφ (I) populations as summarized in Table 1. The histograms show antibody staining (red lines) compared with appropriate isotype staining (blue lines). Flow cytometry plots and histograms as well as associated gated cell percent frequencies are from a representative animal, and the percentage of positive cells is based on the preceding parent populations. Data were collected from a single experiment with n = 3 mice, and 300 000 events were collected by flow cytometry.