In vitro binding assays of HCV domain I nucleotides 1–47 with native and mutant miR-122 species.

(A) Pairing interactions between two miR-122 molecules with 5’ RNA sequences of HCV genotype 1a (nucleotides 1–47). The position of the C3U nucleotide change is indicated. (B) HCV oligonucleotides were incubated with increasing concentrations of native miR-122 or the neuron-specific miR-124, and the resulting complexes were separated by native gel electrophoresis mobility shift assay. The migrations of free miRNA, one miR122-HCV complex (HCV:miR-122) and two miR-122-HCV complexes (HCV-miR-1222) are indicated. (C,D) Wild-type and C3U domain I RNAs were incubated with native miR-122 (C) or mutant miR-122 (D). (E) The seed-match sequence of miR-122 binding site 1 in wild-type and C3U RNAs was mutated and then incubated with increasing amounts of native miR-122. Resulting oligomeric complexes was resolved by non-denaturing gel electrophoresis. The data are representative of three independent replicates. Nucleic acids were visualized after staining with SYBR Gold (Thermo Fisher).