Figure 3.

The eIF2α/ATF4 pathway is involved in the adaptive response in the double injury model. Huh7 cells, expressing empty vector, Core, NS3/4A and NS5A HCV proteins were treated with and without 50 µmol/L menadione. In some experiments, cells were pre-treated with 5 mmol/L NAC 30 min prior to addition of menadione. Protein levels of total- and phosphorylated-eIF2α, glucose-regulated orotein of 78kDa (GRP78) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (A) and ATF4 (B) were determined by Western blotting as described in Materials and Methods. mRNA levels of DDIT3 (C) and HSPA5 (D) were evaluated by quantitative polymerase chain reaction (qPCR). The relative expression was normalized based on the expression of 18S. A t-test was performed to compare the means and the asterisks represent the p value ** ˂0.007 and * ˂0.03. n.s = non-significant, (p values > 0.05 are considered not statistically significant).