Fig. 6. Protein level confirmation of human scRNA-seq observations in large animal comparator.

a, b Protein level confirmation performed using immunofluorescence staining of wounded and untreated (W, left, blue, n = 3 independent wounds) vs. wounded and FAK-inhibited (W_HF, right, red, n = 3 independent wounds) porcine dermal tissue sections at POD7 (from Figs. 1, 2). Staining for YAP (encoded by the gene YAP1; **p = 0.0015), which contributes to myofibroblast differentiation, mechanotransduction, and scar formation, or EGR1 (*p = 0.0415), MFGE8 (*p = 0.0499), and MMP1, which contribute to regenerative healing and collagen degradation. Scale bar: 200 µm. Magnified image scale bar: 50 µm. c, d Western blot protein quantification of human fibroblasts from repeated experiments utilizing our collagen scaffold system of YAP1 (n = 4 independent collagen scaffolds per condition, *p = 0.0493), EGR1 (n = 2 independent collagen scaffolds per condition, *p = 0.0134), MFGE8 (n = 6 independent collagen scaffolds per condition, *p = 0.0164), and MMP1 (n = 4 independent collagen scaffolds per condition, *p = 0.0471). e, f Schematic of the proposed cellular mechanism of action showing how increased mechanical stress drives fibrosis and scar formation, while FAK inhibition unlocks AKT activation of EGR1 and MFGE8. Statistical comparisons were made using either unpaired (b) or paired (d) two-tailed t-tests. Each datapoint represents an independent wound or collagen scaffold. All data represent mean ± SEM. Representative images are shown from similar images across all experiments.