Murine Diaphragm and Heart Cell Isolation

Following euthanasia, diaphragms were dissected from 8–10-month-old WT and mdx mice, washed in 1% Pen/Strep in PBS, and minced. Digestion was completed using Liberase TL (Sigma) for 2 hours at 37 °C on an orbital shaker (20 rpm). Heart: the heart was dissected from 13–16-month-old mdx mice using a Lagendorff-free method^{60 (link)} describe below. Cells were filtered through a 40 μm cell strainer and incubated with antibodies F4/80, Cd11b and DAPI (Thermo Fisher Scientific) for 20 minutes on the benchtop covered from light. Samples were washed and sorted by the Flow Cytometry/Cell Sorting Rutgers Core Facility.