Cav1 tyrosine 14 regulates FAK exchange. (A) Mgat5^+/+ cells were not transfected or transfected for 2 d with either control (CTL) siRNA or Cav1 siRNA and labeled with Hoechst and anti-Cav1 antibodies. Quantification of Cav1 mean intensity relative to Alexa568-phalloidin labeled F-actin (not depicted) is shown as a bar graph (n = 3; ±SEM; *, P < 0.05). (B) Cell lysates from untransfected and CTL and Cav1 siRNA transfected Mgat5^+/+ cells were subjected to Western blot analysis for Cav1 and β-actin. (C) Percent intensity (±SEM) in the bleached zone of FAK-GFP during recovery and quantification of percentage of recovery (box) are shown for Mgat5^+/+ cells transfected with FAK-GFP alone or with FAK-GFP and either CTL or Cav1 siRNA transfected Mgat5^+/+ cells (n = 3, ±SEM; *, P < 0.05). Mgat5^+/+ (D) and Mgat5^−/− (E) cells were cotransfected with FAK-GFP and either Cav1-mRFP or Cav1Y14F-mRFP and subjected to FRAP analysis of FAK-GFP. Percent intensity (±SEM) in the bleached zone of FAK-GFP during recovery and quantification of percentage of recovery (box) are shown (±SEM; *, P < 0.05). (F) Migration of untransfected Mgat5^+/+ and control, Gal-3, and Cav1 siRNA transfected Mgat5^+/+ cells was determined over a 24-h period in serum-free medium on an FN substrate (10 μg/ml) using a wound-healing assay (n = 3; ±SEM; *, P < 0.05).