FIGURE 5.

RanBPM increases the stability of Mgl-1. A, RanBPM increases the expression level of Mgl-1. 293T cells were co-transfected with the constant amount of Myc-Mgl-1 (0.5 μg) together with or without the increasing amounts of Flag-RanBPM (1, 2, 3, and 4 μg). At 48 h after transfection, cell lysates were prepared, and subjected to immunoblotting with the indicated antibodies. B, RanBPM significantly increases the expression level of endogenous Mgl-1. MDCK cells were transfected with or without increasing the amounts of Flag-RanBPM, and were processed for immunoblotting as described above. C, knockdown efficiency of RanBPM-shRNA1 and RanBPM-shRNA2 was checked by Western blot analysis in MDCK cells (upper panel) and its effect on endogenous expression of Mgl-1 (middle panel). D, RanBPM increases the half-life of Mgl-1. 293T cells were transfected with Myc-Mgl-1 alone (0.5 μg) (upper panels) or together with Flag-RanBPM (2.0 μg) (bottom panels). After 48 h of transfection, cells were treated with cycloheximide (100 μg/ml) and harvested at the indicated time periods. Cell lysates were used for immunoblotting with the indicated antibodies.