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. 2012 Jan 3;109(3):941–946. doi: 10.1073/pnas.1112263109

Fig. 3.

Fig. 3.

Stabilization of HCV RNA requires miR-122 binding and is independent of translation. (A) Upper: miR-122 and mutant miR-122p6 guide-strand sequences. Lower: 5′ terminal sequence of HCV (HJ3-5/GLuc2A virus), with S1 and S2 binding sites shown in red. Point mutations (underlined) in the related S1-S2-p6m-GND mutant (11) are shown above. SL-1 and SL-2 are putative stem-loop structures in the 5′ UTR. (B) Northern blots of HJ3-5/GLuc2A-GND and the related S1-S2-p6m-GND mutant RNA after transfection into Huh-7.5 cells with RNA oligoribonucleotides as in Fig. 1B. (C) Putative secondary structure of the HCV 5′ UTR, showing the location of stem-loop IIId and the G(266-8)C IRES mutation that ablates translation. (D) Northern blot showing HCV RNA abundance in MEFs transfected with HCV RNA (H77S/GLuc2A-AAG) containing (Upper) the WT 5′ UTR vs. (Lower) the translationally inactive G(266-8)C mutant, and supplemented with the indicated miRNAs. (E) PhosphorImager quantitation of HCV RNA in Northern blots of 6-h cell lysates from two independent experiments carried out as shown in D.