microRNA-122 Dependent Binding of Ago2 Protein to Hepatitis C Virus RNA Is Associated with Enhanced RNA Stability and Translation Stimulation
Figure 3
Ago2 protein does essentially not interact with the HCV 3′-UTR.
(A-C) Analysis of 32P-labelled HCV 3′-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. S1B. (B) Analysis of the HCV 3′-UTR RNA after immunoprecipitation with Ago2 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-PTB 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 Ago2 in an immunoblot.
