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. 2018 Apr 17;46(10):5139–5158. doi: 10.1093/nar/gky273

Figure 3.

Figure 3.

miR-122 binding does not shield the 5′ terminus of HCV RNA against PKR recognition. Huh-7.5 cells were electroporated with siPKR or siControl (siCon) at day –3 and at day 0 cells were electroporated again with the indicated siRNA, and either (A) wild-type or GNN FL HCV RNA with a firefly luciferase mRNA, or (B) S1+S2p3 SGR or S1+S2p3 GND SGR, a Renilla luciferase control mRNA, and miR-122p3 (miR-122-dependent) or miCon (miR-122-independent). Replication was measured by evaluating luciferase production at the indicated timepoints. (C) Western blot showing knockdown efficiency with antibodies against PKR and β-actin. Percent knockdown ± standard deviation relative to siCon is indicated. (D) Northern blot analysis of FL HCV RNA accumulation during PKR knockdown at day 3. (E) Densitometry quantification of northern blot analysis in (D) normalized to siCon. All data are representative of at least three independent experiments and statistical significance was determined by paired parametric t test.