Figure 2. FBXL19 regulates RhoA stability.

A. MLE12 cells were transfected with three distinct FBXL19 shRNA plasmids (#1 - #3) for 72 h. RhoA, FBXL19, and β-actin expression were analyzed by immunoblotting. B. MLE12 cells were co-transfected with FBXL19-V5 plasmid and RhoA-V5 or RhoAV14-V5, or RhoAN19-V5 plasmids. Cell lysates were analyzed for RhoA-V5 or RhoAV14-V5, RhoAN19-V5, FBXL19-V5, and β-actin by immunoblotting with V5 tag and β-actin antibodies. C. RhoAN19-V5 over-expressed MLE12 cells were co-transfected with vector alone, shFBXL19, or shFBXL19 + FBXL19-V5 plasmids and then cells were treated with CHX (20 μg/ml) for 0 - 4 h. Cell lysates were analyzed for RhoAN19-V5, FBXL19-V5, FBXL19, and β-actin by immunoblotting with V5 tag, FBXL19, and β-actin antibodies. Shown are representative blots from three independent experiments.