Effects of Rab27a depletion on HCV RNA abundance are independent of the 5’ terminal miR-122:HCV complex.

<p>(A) I, Diagram representing interactions between endogenous wildtype (top) and mutant, p3-loop miR-122 molecules (bottom) with a mutant HCV RNA genome (HCV G₂₇G₄₂). Mutations in the HCV G₂₇G₄₂ RNA genome are highlighted in red. A complementary mutation in miR-122 at position 3 is highlighted in red (p3-loop). Additional mutations in p3-loop mimetics at position 9–13 and 18 are highlighted in orange (bottom). II, Diagram representing interactions between endogenous wildtype (top) and p2-8 miR-122 molecules (bottom) with a HCV G₂₇G₄₂ RNA genome. Seed mutations in mutation p2-8 are highlighted in blue. (B) Accumulation of HCV, Rab27a and actin RNA transcripts. Cells were transfected with control or Rab27a siRNAs at day 1, transfected with the p3-loop or p2-8 miR-122 at day 2, and electroporated with the HCV G₂₇G₄₂ RNA at day 3. The cells were transfected again with siRNAs and miR-122 duplexes at day 4. Cells were harvested at day 3 post-electroporation, and RNAs were measured by Northern blot analysis. (C) Quantitation of normalized HCV G₂₇G₄₂ RNA, p3-loop miR-122 and endogenous miR-122 abundances in Rab27a-depleted cells of samples shown in Fig 9B, lane 2 and 3 (marked with triangles). Data in control siRNA treated cells was set as 100%. The data are representative of five independent replicates (*P<0.05 and **P<0.01, Student’s t-test).