Pacific blue conjugated anti mouse cd45

At indicated time points after cardiotoxin injury, TA muscles were dissected, minced, and digested with STEMxyme2 (Worthington Biochemical Corp., Lakewood, NJ, USA). Cell suspension was then serially filtered through 70 and 40 μm nylon meshes (BD Falcon, Franklin Lakes, NJ, USA). All FACS analyses were performed at the Dana-Farber Cancer Institute flow cytometry facilities with a BD FACSAria II SORP UV sorter. Flowjo software was used to analyze the FACS data. Antibodies used include Pacific Blue conjugated anti-mouse CD45 (Biolegend, 30F-11), Alexa 647 conjugated anti-mouse CD68 (Biolegend, San Diego, CA, USA, FA-11), and Brilliant Violet 605 anti-mouse CD206 Antibody (Biolegend, C068C2). For MACS, cells were incubated with FITC-labeled F4/80 and CD11b antibody (eBiosciences) for 30 min in 4 °C and magnetically separated using an EasySep Mouse FITC Positive Selection Kit (Stemcell Technology, Vancouver, BC, Canada). Cell numbers were determined using Countess automated cell counter (Invitrogen).