Fitc conjugated anti cd31

TA, GC, and Qu muscles were used in this study. Mononuclear cells from uninjured or injured limb muscles were prepared using 0.2% collagenase type II (Worthington Biochemical) as previously described [29 (link)]. FITC-conjugated anti-CD31, -CD45, PE-conjugated anti-Sca-1, and biotinylated-SM/C-2.6 [30 (link)] antibodies were used for satellite cell staining.
For detection of macrophages or neutrophils, FITC-conjugated anti-CD45 and PE-conjugated anti-F4/80 (Clone; BM8, BioLegend) or PE-conjugated anti-CD11b (Clone; M1/70, BD Pharmingen), APC-conjugated anti-Ly6G (Clone; 1A8, BioLegend), and V450-conjugated anti-Ly6C (Clone; AL-21, BD Pharmingen) antibodies were used, respectively. For detection of mesenchymal progenitors, FITC-conjugated anti-CD31, -CD45, PE-conjugated anti-Sca-1, and biotinylated anti-PDGFRα (R&D Systems, Minneapolis, MN, USA) were used as described previously [16 (link)]. Cell sorting was performed using an FACS Aria II flow cytometer (BD Immunocytometry Systems).

Mouse OM-MSCs were obtained from turbinal of wild-type or GFP transgenic mice as previously described [17 ]. After culture expansion in vitro, the isolated OM-MSCs were stained with anti-CD45 (BD, 553079), anti-CD34 (BD, 560238), FITC-conjugated anti-CD31 (R&D, FAB3628G), PE-conjugated anti-CD73 (BD, 550741), anti-CD105 (BD, 562761), and APC-conjugated anti-CD90 (R&D, FAB7335R). Isotype control was used to determine cell gating. One hundred thousand cells/sample were collected using FACS Canto II (BD). We analyzed data using FlowJo V10.5.3 software.