FIGURE 2:

N-glycosylation is not required for JAM-A transport but regulates homodimerization and stability. (A) CHO cells expressing wt or N185Q human JAM-A were incubated with sulfo-NHS-biotin to label surface-exposed proteins. Biotinylated proteins were precipitated using streptavidin–agarose and detected by Western blot analysis. The data are representative of three separate experiments. (B) CHO cells expressing wt, N185Q, or 6163 JAM-A and some cells were incubated with BS³ to cross-link proteins. Cells were processed for Western blot analysis. (C) CHO cells expressing wt or N185Q human JAM-A were incubated with 100 μM cycloheximide (CHX) for up to 24 h. Lysates were subjected to Western blot analysis for JAM-A and β-actin and densitometric analysis conducted using ImageJ. Statistical comparisons were assessed between the samples at each time point using a two-tailed Student’s t test. *p < 0.05 between the samples from four separate experiments.