Mdx Mice Treatment with Spironolactone and Prednisolone

Mdx mice were treated using water bottles containing 250 mg/L spironolactone (Sigma-Aldrich, S3378) or 6.7 mg/L prednisolone (Sigma-Aldrich, P6004), the active metabolite of prednisone dissolved in MediDrop containing sucralose (ClearH2O, 75-01-1001). Mice were treated during the peak of inflammation for 10 days for cytokine- and chemokine-level analysis to allow detection of differences in protein expression (4–5.5 weeks of age), for 7 days (3.5–4.5 weeks of age) for flow cytometry, RNA-Seq, and diaphragm IHC or for 14 days (3.5–5.5 weeks of age) for quadriceps IHC and histology. Approximate dosages for spironolactone and prednisolone treatment were 37.5 and 1 mg/kg × day, respectively (8 (link), 33 (link)). The mdx mice given MediDrop vehicle were used as controls for all experiments. Mice were housed 5 per cage and euthanized via cervical dislocation.