S-adenosyl-L-methionine modifies antioxidant-enzymes, glutathione-biosynthesis and methionine adenosyltransferases-1/2 in hepatitis C virus-expressing cells

Figure 5 Effect of S-adenosyl-L-methionine on methionine adenosyltransferase 1/2 turnover. A: Assesment of MAT-mRNA: Huh7 HCV replicon cells (2 × 10⁵ cells) were incubated with 1 mmol/L SAM and then cells were harvested at 24, 48 and 72 h and MAT1 and MAT2 mRNA levels were quantified by real-time RT-PCR and normalized with GAPDH and RPS18 using delta delta Ct method. Mean results from three independent experiments are shown. ^aP≤ 0.05 comparison of SAM vs untreated cells; B: MAT1A/MAT2A Turnover: Measured MAT1A/MAT2A expression represented as a total; C: MAT1 and MAT2 protein expression: Huh7 HCV replicon cells (2 × 10⁵ cells) were incubated with SAM 1 mmol/L. Cells were harvested at 24, 48 and 72 h and total proteins were obtained. Cell lysates were prepared, and equal amounts of protein extracts (40 μg) were subjected to immunoblot analysis to detect MAT1, MAT2 and actin levels. SAM: S-adenosyl-L-methionine; HCV: Hepatitis C virus; MAT: Methionine adenosyltransferase.