Fig 1: sTHY1 treatment rescues the myofibroblast-like phenotype of proband fibroblasts.(A) ** Staining of actin filaments (phalloidin) in control and proband fibroblasts with or without sTHY1. (B,C) Quantification of the cell area (B) and mean fluorescence intensity (C; MFI) of phalloidin-stained cells with or without sTHY1. Proband cells display increased cell size and phalloidin intensity, which is rescued with THY1 treatment. The control and proband phalloidin data without sTHY1 are the same data presented in Figure 3C,D. (D) Immunostaining of PDGFRβ ** in control and proband fibroblasts with or without sTHY1. (E) ** Quantification of the MFI of PDGFRβ-stained cells with or without sTHY1. ** Proband cells display increased PDGFRβ expression, which is rescued with Thy1 treatment. (F)pH3 Immunostaining (proliferation marker) in control and proband fibroblasts with or without sTHY1. (G) Quantification of the percentage of pH3+ nuclei in cells with or without sTHY1. Proband cells display significantly fewer pH3+ nuclei. sTHY1 induces a modest increase in proliferation in control cells but significantly rescued the impaired proliferation in proband cells back to levels comparable to the control. All quantifications are reported as Fold Change compared to control. (H-I) Quantitative real time PCR (qPCR) analysis of TGFB1 (H) and ELN(I) in control and proband fibroblasts with or without sTHY1. Proband cells display increased TGFB1 and ELN expression, which is reversed after sTHY1 treatment. Cells were treated with sTHY1 (1 μg/mL) or vehicle control for 48 hours in the presence of growth serum for phalloidin, PDGFRβ, pH3, and qPCR analysis. Statistics for panels H and I were calculated using unpaired Student’s t-tests. Statistics for panels B, C, and G were calculated using two-way ANOVAs with Šídák’s multiple comparisons test. Statistics for panel E were calculated using a two-way repeated measures ANOVA with Šídák’s multiple comparisons test. Each data point represents an independent experiment. Data are presented as the mean ± S.D. Scale bars: 50 μm (A), 100 μm (D,F).