FIGURE 3.

Cdc42 and Tuba were significantly knocked down using shRNA. Lentivirus-encoding shRNA targeting Cdc42 (A) and Tuba (B) were used to infect MDCK cells (24). The lentiviral construct contained a GFP, which allowed us to use FACS to generate single cell clones. Stable knockdown cell lines were then grown, and Western blotting was performed to determine efficacy of knockdown. By Western blotting, there was complete knockdown of Cdc42 (A) and 70–80% knockdown of Tuba (B). Western blotting of GAPDH is shown to demonstrate equal protein loading in the gel lanes.