Figure 5

Overexpression of Grp78 decreases Rock activity in hepatocellular carcinoma cells. (A), lysates from mock- and Grp78 transfected cells were precipitated using Rhotekin-RBD bound sepharose beads and blotted with anti-RhoA. Corresponding total RhoA is shown in lower blot. (B), A GST pulldown assay using lysates from Grp78 siRNA and control siRNA transfected Grp78 overexpressing cells, indicating RhoA-GTP level. (C), Grp78 knockdown in Rock dominant negative recombinant (KDIA) transfected cells caused obvious stress fiber formation, indicating endogenous Rock activation. (Scale bar = 10 μm). (a), mock-transfected cells.(b) KDIA transfected cells. (c) Control siRNA transfected KDIA cells. (d), Grp78 siRNA knockdown KDIA cells. (D), Representative image of a transwell assay using Grp78 siRNA and control siRNA transfected KDIA cells, indicating Grp78 knockdown rescued the invasion of KDIA cells. The experiments were repeated for three times in triplicate.