FIGURE 5.

BAR modulates BI-1 protein stability. A, 293T cells were co-transfected with BI-1-HA and pcDNA3 vector (Vector), Myc-BAR (BAR), or Myc-BARΔRING (BARΔRING) for 24 h. Cells were then treated with cycloheximide (CHX, 20 μg/ml) for the indicated times to inhibit protein synthesis. Cell lysates were analyzed by immunoblotting using mouse anti-HA and rabbit anti-BAR antibodies. α-Tubulin served as a loading control within each group. The amount of total protein used in each group was adjusted and at time 0, a similar densitometric signal was achieved. B, percentage of the remaining BI-1 protein at each time point after inhibiting protein synthesis using cycloheximide in A was quantified by densitometry (mean ± S.E.; n = 2).