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. Author manuscript; available in PMC: 2017 Aug 10.
Published in final edited form as: Nat Med. 2016 Nov 14;22(12):1475–1481. doi: 10.1038/nm.4211

Figure 1.

Figure 1

Inhibition of miR-208b and miR-499a-5p during HCV infection improves hepatocyte response to IFN-β by increasing IFNAR1. (a) Expression of MYH7 and MYH7B in Huh7 cells infected with HCV MOI 1.0 for 48 h. (b) Expression of ISGs MX1, OAS1 and PKR in HCV-infected Huh7 cells transfected with inhibitors against miR-208b and miR-499a-5p (myomiR inh) or non-targeting control inhibitor (NC) for 48 h and then treated with IFN-β for 6 h. (c) HCV viral titer and copy number from samples in b. (d,e) RNA hybrid analysis was used to show miRNA–target interactions for miR-208b and miR-499a-5p, which bind to numerous sites of the IFNAR1 3′ UTR. (d) Schematic of miR-208b and miR-499a-5p binding sites on the IFNAR1 3′ UTR. p(A)n, poly(A) site. (e) Sequence alignment of miR-208b and miR-499a-5p and their binding sites in the IFNAR1 3′ UTR; miRNA–target seed regions are underlined and nucleotides for IFNAR1 3′ UTR are indicated. (f) Levels of IFNAR1 and IFNAR2 mRNA and (g) IFNAR1 surface protein expression in HCV-infected Huh7 cells transfected with myomiR inh or NC inh. Gene expression for a–c,f was assessed using qPCR, and the reference sample for relative quantification was (a–c) mock-infected cells transfected with myomiR inhibitors or NC inhibitor assayed at 48 h or (f) HCV-infected cells transfected with NC inhibitor assayed at 48 h. Data are from one experiment representative of three or more experiments (a,b,f, mean ± s.e.m.; c,g, box and whisker plots show median values (line), 50th percentile values (box outline) and minimum and maximum values (whiskers)). Unpaired Student’s t-test was used for all statistical comparisons; *P < 0.05, **P < 0.01, ***P < 0.001.