Inhibition of miR122 function either by VV infection or by I7 protease expression.

<p>(A) HCV replicon cells were mock-infected (lane 2) or infected with VV in M.O.I. = 1 (lane 3), 5 (lane 4), 10 (lanes 5 and 6). 100 ug araC was also added (lane 6). Twenty-four hrs later, cell lysates were analyzed by SDS-PAGE and Western blotting with antibodies against Dicer protein (upper panel), NS5A and NPT proteins for the expression of HCV subgenomic RNAs, or Erk2 protein as the loading control. Cell lysates from mock-infected HuH7 cells (lane 1) were served as a negative control for the detection of NS5A and NPT. (B) HCV replicon cells were mock-transfected or transfected with the indicated amount of the plasmid expressing I7 protease (lanes 1–6). Thirty-two hrs after transfection, 10 uM MG132 was also added (lanes 5 and 6). Sixteen hrs later, cell lysates were analyzed by SDS-PAGE and Western blotting with antibodies against Dicer protein, NS5A protein for the expression of HCV subgenomic RNAs, NPT protein (the thin arrow marks the position of NPT protein expressed from the transfected plasmids while the thick arrow marks the position of core-NPT fusion protein encoding from HCV subgenomic RNAs.), V5 tag to detect the expression of I7 (The arrow marks the position of I7 in lane 6.) or Erk2 protein as the loading control.