Figure 2
Screening of the effect of GRP78-shRNAs and the establishment of cell clones that stably expressing GRP78-shRNA. (A) Fluorescence observation of the transfection efficiencies of shGRP78s in SMMC7721 cells. ShGRP78s containing GFP tag were introduced into SMMC7721 cells as described under “materials and methods”. After 72 h, GFP fluorescence were observed by inverted fluorescent microscope(scale bar:25 μm). (B) Western blot analysis of GRP78 levels in GRP78-shRNAs transiently transfected cells. The GRP78 levels were presented as the ratio of GRP78 to β-actin. (C) Western blot analysis of GRP78 levels in cells that stably expressing shGRP78-3. The contents of GRP78 were expressed as the ratio of GRP78 toβ-actin. (D) Western blot analysis of GRP94 levels in clone C3 and C4 that stably expressing shGRP-3. The contents of GRP94 were expressed as the ratio of GRP94 to β-actin. All the experiments were repeated for three times, the values were presented as ± SE and analyzed by One-Way ANOVA (Columns,mean of three separate experiments; bars, SE; *, values significantly different at the 5% levels).