Cell-surface expression of mLMIR5. (A) Ba/F3 cells transduced with a Flag-tagged mLMIR5 or mock were stained with FITC-conjugated mouse IgG1 or anti-Flag Ab as well as polyclonal goat IgG or anti-mLMIR5 Ab, followed by PE-conjugated anti-goat IgG F(ab′)₂. (B) Analysis of mLMIR5 expression on hematopoietic cells derived from C57BL/6 mice. Single-cell suspensions were prepared from BM, PB, peritoneal cavity, and thymus. Cells were stained with control IgG or anti-mLMIR5 Ab followed by PE-conjugated anti-goat IgG F(ab′)₂ and FITC-conjugated mAbs as indicated. In BM, PB, and peritoneal cells, FSC^highSSC^high populations representing myeloid lineage were gated and analyzed for mLMIR5 expression. In thymus, the FSC^lowSSC^low populations representing lymphoid lineage were analyzed. (C) Single-cell suspensions were prepared from spleen. After B220⁺, CD3⁺, CD11b⁺, or CD11c⁺ cells were sorted by using FITC-conjugated Abs, these cells were stained as described in panel B. (D) Analysis of mLMIR5 expression on murine BM-derived cells. BMMCs, BMmDCs, BMpDCs, and BMMΦ were stained with control IgG or anti-mLMIR5 Ab followed by PE-conjugated anti-goat IgG F(ab′)₂. The result of control or mLMIR5 staining is shown as a filled or bold-lined histogram, respectively. All the data are representative of 3 independent experiments.