Rho/ROCK Inhibition Promotes TGF-β3-Induced Tenogenic Differentiation in Mesenchymal Stromal Cells

Figure 2

Fluorescence microscopy in equine monolayer MSC. Cells were incubated with 10 ng/ml TGF-β3 and/or 10 μM ROCK inhibitor Y-27632 and analysed at days 1 and 3. Representative images show phalloidin staining of the actin cytoskeleton, which is disrupted upon ROCK inhibition, nuclear staining with DAPI, and Smad 2/3 immunostaining with nuclear translocation upon TGF-β3 and TGF-β3 plus Y-27632 stimulation. Differences between groups are clearly visible on days 1 and 3. The graphs represent the data from quantitative image analyses in cells from 3 donors, indicating a constant amount of total Smad 2/3 (left) and nuclear translocation upon incubation with TGF-β3 and Y-27632 (right). Bars represent the median values, error bars represent the 95% confidence intervals, and the asterisk marks a difference between the indicated groups with .