Role of PKR in the Inhibition of Proliferation and Translation by Polycystin-1
Figure 3
Effects of PKR siRNA and PKR-K296R on PC1-inhibited proliferation and translation. (a) Effects of PKR siRNA and PKR-K296R on PC1-inhibited proliferation. HEK293T and those stably transfected full-length PC1 were transfected with GFP vector, PKR-siRNA or PKR-K296R, and then plated in multiple wells of a 96-well plate and grown for 24 hr for cell proliferation assays. Cells from the sample preparations were collected for immunoblotting. PC1, GFP-tagged full-length PC1; Ctrl, GFP; K296R, PKR-K296R. Upper panel, averaged data (N=4). Lower panel, effectiveness of PKR-K296R transfection and PKR siRNA assessed by immunoblotting. (b) Effects of PKR-K296R and PKR siRNA on PC1-inhibited translation. HEK293T and those cells stably transfected full-length PC1 were transfected with GFP vector, PKR-siRNA, or PKR-K296R and then for 35S pulse labelling assays, followed by SDS-PAGE and immunoblotting assay with the antibody against P-PKR. β-actin served as loading control. P-PKRs, band with short exposure of 0.5 min; P-, band with common exposure of 2 min.