Figure 2. pUR4 attenuates mouse pathologic myofibroblast phenotype.

(A) pUR4 decreases collagen I staining in CF isolated after cardiac I/R injury by immunofluorescence. Scale bars, 100 μm. (B) Matrix collagen I deposition in activated MF is attenuated upon pUR4 treatment (representative immunoblots, left, and densitometry, right). n=5. (C) Cellular collagen I protein level expression (without matrix) (representative immunoblots, left, and densitometry, right). n=5. (D) Collagen I transcript level expression. n=6. (E) pUR4 decreases cell proliferation monitored by counting cell number with a hemocytometer. n=6. (F) Phospho-histone H3 (pHH3) staining in mouse MF shows a decrease in pHH3⁺ cells in pUR4-treated cells (representative pictures, left, and quantification, right). Scale bars, 100 μm. n=3. (G) RT-qPCR of proliferative-related genes in pUR4 and III-11C-treated mouse MF. n=6. (H) Scratch wound-healing assay in mouse CF cell migration; pictures were taken at 0 h, 12 h and 24 h post-scratch. Black dotted lines denote the wound borders (representative photographs, left, and mobility quantification, right). Scale bars, 1000 μm. n=6. (I) Increased β1 integrin internalization was found in MF treated with pUR4 compared to III-11C peptide (representative immunoblots, left, and densitometry, right). n=5. (J) Expression of phosphorylated focal adhesion kinase (p-FAK) was evaluated by Western blotting and quantified relative to total FAK expression (representative immunoblots, left, and densitometry, right). n=4. (K) Alpha-smooth muscle actin (α-SMA) staining in III-11C and pUR4-treated MF. Scale bars, 100 μm. (L) Vimentin staining in peptide-treated cells. Scale bars, 100 μm. Statistical significance was determined with paired t-test: *P<0.05, **P<0.01, ***P<0.001. Data are represented as mean ± SEM.