FIGURE 3.

GRIM-19 overexpression did not alter STAT3 transcriptional activity. (A) The expression and phosphorylation of STAT3 after transfection with pcDNA3 or pcDNA3_GRIM-19 were evaluated by western blot analysis. β-actin was used as an internal control for loading. (B) To evaluate subcellular localization of phospho-STAT3, STAT3, and GRIM-19, HCVcc-infected Huh7 cells were transfected with pEGFP-C1 or pEGFP-C1_GRIM-19. After 48 h, the cells were subjected to subcellular fractionation followed by western blot analysis. PARP and SOD1 were used as markers for the nucleus and cytosol. (C) The mRNA levels of bcl2 and mmp2 were assessed by RT-PCR with a gene-specific primer set. The mRNA expression was normalized to β-actin as a reference and expressed relative to the density of Huh7 cells. Values represent means ± SD. (D) Examination of the induction of apoptosis by overexpression of GRIM-19 in Huh7 cells or FGR cells. The cells were transfected with pcDNA3 or pcDNA3_GRIM-19. After 48 h, apoptosis was determined by a Annexin V/PI staining.