Fig. 1. Distribution of bone marrow-derived monocytes according to the expression of Ly6C by FACS analysis. (A) Representative FACS plots show the gating strategy used for the identification of murine monocyte subsets isolated from bone marrow based on their relative expression of Ly6C expression by all flow cytometry and cell sorting experiments. In brief, after exclusion of apoptotic cells by DAPI, a lineage negative selection was performed (CD3e, CD19, NK1.1, CD49b, CD117, Ly6G) to deplete mature hematopoietic cells and their committed precursors. The remaining cell fraction was gated for CD11b to include all monocytic cells and analyzed for the expression of Ly6C. (B) The increase in CD11b⁺ monocytes is illustrated for all three genotypes (SRL^-/-, RL^-/- and L^-/-) by plotting the percentage of CD11b positive cells within all living bone marrow cells. (C) Representative FACS dot blots of Ly6C expression on CD11b⁺ monocytic cells derived from bone marrow of SRL^-/-, RL^-/- and L^-/- mice. The percentage of Ly6C^high and Ly6C^low monocyte subsets of each genotype within the depicted gates is given. (D) The distribution of CD11b⁺ monocytes according to their expression of Ly6C illustrates the influence of RAG2 deficiency on Ly6C^high monocyte subset as well as the influence of SOCS-1 deficiency on Ly6C^low monocyte subset. Data are shown as mean ± SEM of 10 mice per group from three independent experiments. Significance was calculated in comparison to all other groups by one-way ANOVA/Tukey's test (*P<0.05, **P<0.01, ***P<0.001).