Figure 6.

Paxilin expression and FAK phosphorylation are negatively regulated by Ski in cardiac myofibroblasts. First passage (P1) cardiac MFB were grown to reach 70% confluency and infected with either Ad‐LacZ or AdSki (MOI of 50, 100, or 150) for 48 h. Untreated controls were serum‐deprived, but not infected with either virus. Expression of exogenous HA‐tagged Ski was confirmed using anti‐HA antibodies and β‐tubulin was used as a loading control. (A) Representative immunoblots of total cell lysates probed for paxillin. (B) Histogram showing data obtained in A. Paxillin protein levels are significantly decreased in response to AdSki infection, as compared to AdLacZ‐infected and uninfected controls. Data shown are representative of the means ± SEM where n = 3; *P ≤ 0.05 for AdSki50 compared to untreated and AdLacZ‐50 controls; ***P ≤ 0.001 for AdSki150 compared to untreated and AdLacZ‐150 controls. C: Representative immunoblots of whole cell lysates probed for immunoreactivity to total FAK and phospho‐FAK (Tyr 397). n.s., non‐specific antibody binding. (D) Histogram showing data obtained in A. FAK phosphorylation at Tyr 397 is significantly decreased with Ad‐Ski infection at 150 MOI, compared to AdLacZ‐50 and uninfected controls. Data are representative of the mean ± SEM where n = 4; *P ≤ 0.05 for AdSki150 compared to controls. (E) Representative immunoblots of total cell lysates probed with vinculin antibody. (F) Histogram showing data obtained in E. No significant differences were observed among treatment groups. Data shown are representative of the means mean ± SEM where n = 8; P = 0.2619. (G) Representative immunoblots of total cell lysates for integrin‐β1 antibody. (H) Histogram showing data obtained in A. Data shown are representative of the mean ± SEM where n = 4; P < 0.6506.