FIGURE 3:

N-glycosylation regulates JAM-A–mediated increases in barrier integrity. (A) CHO cells expressing empty vector or wt or N185Q JAM-A were grown on 0.4-μm Transwell inserts for 48 h. FITC-dextran (10 kDa) was added to the upper chamber and measured from the lower chamber after 2 h of incubation. Data shown are representative of three separate experiments run in triplicate. Statistical differences were determined by one-way analysis of variance (ANOVA) with Tukey’s posttest. *p < 0.05 vs. empty vector and N185Q. (B) The same cells as in A were grown on RTCA plates, and impedance was assessed for 30 h. Data shown are representative of four separate experiments run in quadruplicate. Statistical differences were determined by two-way ANOVA with Bonferroni posttest against empty vector. (C) CHO cells transfected with empty vector or wt or N185Q human JAM-A were assayed for Rap1 activity by pull down using GST-RalGDS-RBD. (D) Quantification. *p < 0.05 vs. EV; ***p < 0.01 vs. EV; ^#p < 0.05 vs. wt by one-way ANOVA with Tukey’s posttest from four separate experiments.