FIGURE 8.
Effect of tsiRNA expression on the hepatocyte metabolism. (A) Northern blot analysis for miR-122 in hepatocytes. Huh7 cells were treated with indicated siRNA or tsiRNA-expression vector. (Empty) pGD vector containing only the U6 promoter and termination sequences without insert for transcription (negative control). On day 2, total RNA samples were loaded onto 15% PAGE/7 M urea and hybridized with 32P-labeled complementary oligonucleotide for detection of miR-122. (Upper) The marker is a radio-labeled 10-nt ladder. (Lower) U6 snRNA was used as an internal loading control on 5% PAGE/7 M urea. (B) IFN response to cellular expressed tsiRNA. poly(I:C) as a strong IFN pathway stimulator was used as a positive control. Total RNA was isolated from cells on day 2 post-transfection and subjected to semiquantitative RT-PCR amplification of mRNAs from genes encoding IFN-β (35 cycles), OAS1 (20 cycles), and MxA (30 cycles). β-Actin mRNA expression was analyzed as an internal control.