Figure 3. fRHOA and fCDC42 temporarily increase cell proliferation of Pggt1b-deficient K-RAS^G12D–expressing fibroblasts.

(A) Western blots showing expression of the Myc tag in fRHOA-transfected fibroblasts and the HA tag in fCDC42-transfected fibroblasts. Identical blots were incubated with antibodies recognizing RHOA and CDC42. Note the slower electrophoretic mobility of the tagged proteins. (B) Proliferation of Cre-adenovirus–treated primary Pggt1b^fl/flK^LSL fibroblasts transiently transfected with empty plasmid (filled squares) or plasmids encoding Myc-tagged fRHOA (open squares), HA-tagged fCDC42 (filled triangles) or both fRHOA and fCDC42 (black circles). Values are the means of triplicate measurements. The experiment was repeated 3 times with similar results. Transfection of wild-type fibroblasts with these plasmids did not affect cell proliferation. (C) Upper panels show photos of Pggt1b^fl/flK^LSL fibroblasts transiently transfected with the plasmid encoding fRHOA and fCDC42 and then treated with the Cre-adenovirus. Eight days later, the cells were treated with vehicle (–FTI) or 10 μM FTI-276 (+FTI) for 72 hours. Lower panels show that cells not treated with the Cre-adenovirus were unaffected by the FTI.