Figure 3: S16D-Hsp20 Cardiomyocyte-Conditioned Medium Activates Myofibroblast Differentiation

Adult mouse cardiac fibroblasts were cultured in conditioned medium from Ad.GFP, Ad.S16D, Ad.S16A. Fibroblast proliferation and markers of myofibroblast differentiation were assessed. (A) Quantification of fibroblast proliferation was determined using the MTT assay. Values are fold changes relative to those of Ad.GFP. Relative gene expression levels are shown for (B) Col1a1; (C) Col3a1; (D) TGFβ1; and (E) IL-6. Values are fold-changes relative to those of Ad.GFP. (F) Representative confocal images (original magnification: ×40) of fibroblasts stained with αSMA (green) plus DAPI to visualize nuclei (blue); scale bar: 50 μm. (G) Quantification of αSMA fluorescence intensity. The number of samples (n) per group is indicated on the bar graphs. SMA = smooth muscle actin; Ad.GFP = adenovirus-infected green fluorescent protein; Ad.S16D = adenovirus-infected phosphorylated Hsp16D; Ad.S16A = adenovirus-infected dephosphorylated Hsp16A; Col1a1 = collagen 1a1; Col3a1 = collagen 3a1; IL = interlukin-6; TGF = transforming growth factor; other abbreviations as in Figure 2.

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