Figure 5. Analysis of FN organization after disruption of Rho activity via addition of C3 transferase and exposure to mechanical stimulation.

Epifluorescent images were imported into ImageJ software and deconvolved to analyze the distribution of unstimulated (A) and mechanically stimulated (equibiaxial stretch (B) shear fluid flow (C) FN across the cell. Representative plots of intensity profiles plotted as a function of distance (measured in pixels) versus intensity (measured in gray-scale values) showed FN distribution for unstimulated cells (D) line 1–2 and (E) line 3–4. FN distribution of unstimulated cells exposed to C3 transferase was found to occur at only localized areas in the cell. Plots of intensity profiles of FN distribution after 24 hours of 10% equibiaxial stretch (F) line 1–2 and (G) line 3–4 revealed FN redistributed around the cell periphery, which was reflected in the distinct fluorescence intensity peaks located at the ends of the lines, indicating the periphery of the cell. Analysis of FN distribution after 24 hours of 6±3 dynes/cm² of fluid flow shear stress, (H) line 1–2 and (I) line 3–4 revealed that the FN was distributed around the cell periphery, which was reflected in the distinct fluorescence intensity peaks located at the ends of the lines indicating the periphery of the cell. (scale bar = 10 μm).