Cd41 mwreg30

All experiments were performed using 8- to 12-week-old C57BL/6 wild-type mice in accordance with UCSC IACUC guidelines. Hematopoietic cells were isolated from BM by crushing murine femurs, tibias, hips, and sternums as previously described [35 (link)]. Stem and progenitor cell fractions were enriched using CD117-coupled magnetic beads (Miltenyi). Cells were stained with unconjugated lineage rat antibodies (CD3, CD4, CD5, CD8, B220, Gr1, Mac1, and Ter119) followed by goat-α-rat PE-Cy5 (Invitrogen). Stem and progenitor cells were isolated using fluorescently labeled or biotinylated antibodies for the following antigens: cKit (2B8, Biolegend), Sca1 (D7, Biolegend), Slamf1(CD150) (TC15-12F12.2, Biolegend), CD41(MWReg30, Biolegend), and CD71(RI7217, Biolegend). Cells were sorted using a FACS Aria II (BD Bioscience). HSCs were defined as cKit⁺ Lin⁻ Sca1⁺ Flk2⁻ and Slamf1⁺; MkPs as cKit⁺Lin⁻Sca1⁻Slamf1⁻CD41⁺. Unipotent lineage cells were isolated by the following markers and as described previously [15 (link), 29 (link)]: EPs, Lin(CD3, CD4, CD5, CD8, B220, Gr1, and Mac1)⁻ CD71⁺Ter119^±; GMs, Lin(CD3, CD4, CD5, CD8, B220, and Ter119)⁻ Gr1⁺Mac1⁺ (“GM” cells were positive for both Gr1 and Mac1); T cells, Lin(CD5, B220, Gr1, Mac1, and Ter119)⁻ CD25⁻CD3⁺CD4^±CD8^±; B cells, Lin(CD3, CD4, CD8, Gr1, Mac1, and Ter119)⁻CD43⁻B220⁺.