Analysis of Ly6C⁺ cells in OPN^−/−mdx infiltrates. (A) Muscle leukocytes were isolated from OPN^−/−mdx and OPN^+/+mdx muscles, stained for Gr1 and F4/80, and assessed by flow cytometry. FACS plots show F4/80 plotted against Gr1. Cells enclosed by the blue square represent neutrophils. The frequency of neutrophils is indicated in the top right of each quadrant. (B) Analysis of eosinophils was performed by Siglec F staining on sorted F4/80⁺ cells from OPN^+/+mdx and OPN^−/−mdx muscles. Circled regions in the FACS plots represent the eosinophil population, and numbers indicate eosinophil frequencies (top). Bottom graphs show the quantification of F4/80⁺ cell frequency in the total intramuscular leukocyte population (bottom left) and eosinophils in the intramuscular F4/80⁺ cell population (bottom right). Black bars represent OPN^+/+mdx, and white bars represent OPN^−/−mdx. Animal numbers are indicated on graphs. P-values were generated by Student’s t test. Vertical lines represent SEM.