microRNA-122 Dependent Binding of Ago2 Protein to Hepatitis C Virus RNA Is Associated with Enhanced RNA Stability and Translation Stimulation
Figure 4
Ago1 protein interacts weakly with the HCV 5′-UTR.
(A–C) Analysis of 32P-labelled HCV 5′-UTR RNA after transfection into HeLa cells in the presence or absence of miR-122 or control miR-124 duplexes as indicated. The cells were lysed 6 h after transfection. (A) Aliquots of the cell lysate were used for RNA re-extraction to check the integrity of the input RNA prior to immunoprecipitation. The re-extracted radioactive HCV RNA was visualized after gel electrophoresis and autoradiography. A quantification of the RNA amounts in lanes 2–5 is shown in Fig. S1C. (B) Analysis of the HCV 5′-UTR RNA after immunoprecipitation with Ago1 antibodies. From the precipitate, RNA was re-extracted, and the radiolabelled HCV RNA was visualized after gel electrophoresis and autoradiography. In lane 1, the input RNA is shown (1∶100 dilution). In lane 2, an anti-eIF3 antibody was used as a positive control, and in lane 3, an anti-Flag antibody was used as a negative control. (C) Aliquots of the immunoprecipitates used in lanes 2–5 in (B) were checked for Ago1 in an immunoblot.
