Figure 10.

Changes of cellular localization of Cas in double-mutant Cas-transfected Rat-1 cells. Rat-1 cells were transiently transfected with pFLAG-CMV-5c containing double-mutant (D⁴¹⁶E and D⁷⁴⁸E) or wild-type Cas DNAs and incubated with 40 μM etoposide for 18 h (A– D) or 24 h (E and F). Cells were double immunostained with anti-FLAG M2 mAb (A, C, and E) and paxillin Ab (B, D, and F). Positive staining with anti-FLAG M2 mAb indicates cells expressing double-mutant or wild-type Cas. The images in A, C, and E depict alternate staining of the same cells shown in B, D, and F, respectively. Focal adhesions at the cell bottom are indicated by white arrows. Note that double-mutant Cas could attenuate the Cas degradation and consequently partially blocked the relocalization of focal adhesion proteins into the periphery of cells. Bar, 10 μm.