Apc cy7 conjugated anti mouse ter119

Phenotypically distinct muscle and tumor cell subsets were sorted according to protocols that were previously established to isolate functionally discrete subsets of myofiber-associated cells [10 (link), 11 (link), 1 (link)]. In brief, cells were suspended in HBSS supplemented with 2% FBS. Antibody staining was performed for 20 minutes on ice. The following primary and secondary antibodies were used: APC-CY7-conjugated anti-mouse CD11b (1 in 200, BD Pharmingen, 557657), APC-CY7-conjugated anti-mouse CD45 (1 in 200, eBioscience, 557659), APC-CY7-conjugated anti-mouse TER119 (1 in 200, BioLegend, 116223), APC-conjugated anti-mouse Sca1 (1 in 200, eBioscience, 17-5981-82), PE-conjugated anti-mouse/ rat CD29 (1 in 400, BioLegend, 102207), biotin rat anti-mouse CD184 (1 in 100, BD Pharmingen, 551968), PECY7-conjugated Streptavidin (1 in 200, eBioscience, 25-4317-82). Antibody staining was performed for 20 minutes on ice. Prior to FACS sorting, cells were suspended in 1µg/ml propidium iodide and 10µM calcein blue (Invitrogen) to identify viable cells (Pi⁻Ca⁺). Cells were sorted twice to maximize purity.