FIGURE 5.

The effect of RNA-mediated interference-mediated silencing of HNF-4α on PED/PEA-15 gene expression in HepG2 cells. HepG2 cells were transiently transfected with two HNF-4α-specific shRNA clones (Cl.a, Cl.b). 48 h upon transfection total RNAs were extracted form transfected and nontransfected cells (NT). HNF-4α (A) and PED/PEA-15 (B) mRNA levels were then quantitated by RT-PCR. Bars represent values normalized for β-ACTIN mRNA. C, HepG2 cells were transiently transfected with the HNF-4α shRNA (Cl.b). After 48 h transfected and non-transfected cells were solubilized, and lysates were Western-blotted with antibodies toward HNF-4α, PED/PEA-15, or β-actin. Filters were revealed by ECL and autoradiography followed by densitometric analysis of the autoradiographs. Bars represent the means ± S.D. of four (A and B) and three (C) independent experiments. Asterisks denote statistically significant differences. A representative blot is shown in the left panel of C.