Figure 3.

Wild-type and Rho GTPase-mutant pericyte contractility is dose dependently reversible by small molecule inhibition of Rho kinase. A: Bovine retinal pericytes were transduced with adenoviruses containing dominant-active Rho GTPase (RhoDA), dominant-negative Rho GTPase (RhoDN), transactivator alone (Tet), or mock-infected (control) as above. After 24 hours of culture, cells were trypsinized and replated onto plasma glow discharge-prepared, type I collagen-coated silicon substrates in media containing 0 to 10 μmol/L of the small molecule Rho kinase inhibitor Y-27632, and monitored by real-time phase-contrast imaging. Representative phase-contrast images of inhibitor-treated wild-type, mock-infected pericytes are provided as labeled, where arrows indicate substrate-wrinkling, actively contractile pericytes. Original magnifications, ×400. B: At 24 hours, contractility was assessed by the number of pericytes producing visible substrate wrinkling per each condition, expressed as mean percentages ± SE (n > 100 cells per condition, triplicate experiments.