(a)
(b)

Figure 2: Effects of PKR/eIF2α overexpression on PC1-5TMC-inhibited proliferation and translation in HeLa cells. (a) Effects of eIF2α overexpression on PC1-5TMC-inhibited proliferation. Transfected with GFP or PC1-5TMC, HeLa cells were cotransfected with eIF2α. They were then plated in multiple wells of a 96-well plate and grown for 24 hr for cell proliferation assay; cells from the sample preparations were collected for immunoblotting. 5TMC, GFP-tagged PC1-5TMC; eIF2α, WT eIF2α; Ctrl, GFP. Upper panel, averaged data (N=4,p< 0.05, p< 0.01). Lower panel, effectiveness of transfection of eIF2α assessed by immunoblotting. (b) Effects of PKR overexpression on PC1-5TMC-inhibited protein synthesis. After transfected with GFP or PC1-5TMC, HeLa cells were cotransfected with WT PKR. They were then used for 35S pulse labeling assays followed by SDS-PAGE and immunoblotting assays with the antibodies against P-eIF2α, total eIF2α, and P-PKR. β-actin served as loading control.