Analysis of BM cells and BMMCs derived from DAP10^−/−, DAP12^−/−, and FcRγ^−/− mice. (A,B) Surface expression levels of endogenous mLMIR5 on BM cells and BMMCs derived from WT, DAP10^−/−, DAP12^−/−, or FcRγ^−/− mice were analyzed as described in Figure 2. The mean fluorescent intensity (MFI) of mLMIR5 expression was indicated in BMMCs. (C) WT, DAP10^−/−, DAP12^−/−, or FcRγ^−/− BMMCs transduced with Flag-tagged mLMIR5 were stained with control IgG or anti-Flag mAb followed by FITC-conjugated anti-mouse Ig Ab to confirm transduced-mLMIR5 expression levels (bottom row). Phenotypical analysis of BMMCs was performed as described in materials and methods (top row). (D) Either WT, DAP10^−/−, DAP12^−/−, or FcRγ^−/− BMMCs transduced with mLMIR5 were stimulated with control IgG or anti-mLMIR5 Ab. The amount of phosphorylated ERK1/2 was analyzed as described in Figure 4A. Vertical lines have been inserted to indicate a repositioned gel lane. (E) Either WT, DAP10^−/−, DAP12^−/−, or FcRγ^−/− BMMCs transduced with mLMIR5 were stimulated on FN-coated plates. Percentages of adherent cells were estimated. All data points correspond to the mean and the SD of 3 independent experiments as indicated. (F) Relative expression levels of DAP10, DAP12, and FcRγ among WT, DAP10^−/−, DAP12^−/−, and FcRγ^−/− BM or BMMCs were estimated by using real-time PCR as described in “Gene expression analysis.” The amount of expression was indicated relative to that in wild-type BM or BMMCs. Data are representative of 3 independent experiments.