(A) Experiment schematic whereby a cohort of mice received bilateral VML defects to the tibialis anterior muscles followed by intramuscular injection of vehicle (PBS) or TGFBR2-inhibitor ITD1. Spatial transcriptomics analysis was performed at 7 dpi. (B) Integration of spatial transcriptomics data sets with matched, cell type–annotated scRNA-Seq data sets using Seurat label transfer. Plots are annotated with P values. n = 3–4 tissues from 3 male and 3 female mice. Statistics performed using 2-sided, 2-sample t test. Color bars indicate Seurat prediction scores. P < 0.05 were considered significant. (C) (Left) Representative immunohistological stains of macrophages (CD68), PDGFRa⁺ mesenchymal cells, and RFP⁺ MuSCs and their progeny from tissues treated with ITD1. Scale bars: 500 μm. (Right) Quantifications of immunohistological images. *P < 0.05, **P < 0.01. n = 3–4 tissues from 3 male and 3 female mice. Statistics performed using 2-sided, 2-sample t test. (D) Volcano plots showing differential gene expression as a result of ITD1 treatment in the defect (left) and transition zones (right). Differential expression was calculated using MAST, and genes with a adjusted P < 0.05 were considered significant (yellow). Green indicates log₂ fold change greater than 0.0585 and P > 0.05. (E) Violin plots of gene module scores for the macrophage, mesenchymal-derived cell predicted to be involved in active signaling pathways after VML. *P < 0.05, ^#P = 0.06, by 2-sample, 2-sided t test comparing average module scores for each tissue in each zone across treatments. n = 3–4 tissues per group from 2 male and 2 female mice. Magnification, 20×.