FIGURE 6.
Expression of vinculin Y1065F mutant protein decreases the interaction between vinculin and talin in tracheal smooth muscle tissues. Talin was immunoprecipitated (IP) from tissues expressing WT vinculin, vinculin Y1065E, vinculin Y1065F, or sham-treated tissues after 5 min of stimulation with 10−4 m ACh or without stimulation (US). A, representative immunoblots against talin, vinculin and EGFP from muscle extracts from a single experiment. Endogenous metavinculin and EGFP vinculin both migrate at about 145 kDa, whereas endogenous vinculin migrates at 116 kDa. B, relative levels of vinculin that co-precipitated with talin for each treatment group. In ACh-stimulated sham-treated tissues and tissues expressing WT vinculin or vinculin Y1065E more vinculin co-precipitated with talin than in unstimulated tissues. In Y1065F-treated tissues stimulated with ACh, the amount of vinculin that co-precipitated with talin in response to ACh did not increase significantly. *, significant difference between ACh and unstimulated tissues (n = 4, p < 0.05). Error bars, S.E.