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Fig. 5. Decreased ezrin threonine phosphorylation in Foxj1-null airway epithelial cells. (A) Wild-type (+/+) and Foxj1-null (–/–) primary MTE cells cultured as in Fig. 2 and analyzed at ALI day 14 were immediately fixed in TCA, calycA or control buffer then 5 µg of protein was subjected to immunoblot analysis with phosphothreonine (P-T) ERM-specific and other indicated antibodies. Phosphothreonine ezrin (P-T-ezrin) and moesin (P-T-moe) are indicated by arrows. Ezrin (clone 3C12) and moesin (Upstate Biotech) were detected in whole cell lysates. (B) P-T-ezrin and P-T-moesin expressed as a ratio of ß-catenin (ß-cat) determined by densitometry from immunoblots as in A. Data represents the mean and standard deviation of expression from four independent preparations. The asterisk indicates a difference in means (P<0.05). (C) Expression of moesin in the detergent-insoluble fraction of primary culture MTE cells from wild-type or Foxj1-null cells from ALI day 14. Fractions were prepared and 5 µg of protein per lane were analyzed as in Fig. 2C.
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