Scrambled mirna

The miRNA mimics, siRNA, and scrambled miRNA used as a negative control were purchased from Genolution Pharmaceuticals (Seoul, South Korea). The sequences are as follows: scrambled control sense, 5′-UUUUAACUCAGUAUUUUUA-3′ and antisense, 5′-UAAAAAUACUGAGUUAAAA-3′; Prox1 siRNA sense, 5′- GAGUUGACAUUGGAGUGAA-3′ and antisense, 5′- UUCACUCC AAUGUCAACUC-3′. The LNA™ microRNA Power Inhibitor for hsa-miR-466 and the negative control inhibitor (NC inhibitor) were purchased from Exiqon (Vedbaek, Denmark). The sequences are as follows: inhibitor for hsa-miR-466, 5′-GTGTTGCGTGTATGTGTA-3′; NC inhibitor, 5′-GTGTAACACGTCTATACGCCCA-3′.

Mouse monoclonal anti-HCV core antibody and rapamycin were purchased from Thermo Fisher Scientific (Rockford, IL, USA). Mouse monoclonal anti-SREBP-1c, mouse monoclonal anti-fatty acid synthase (FAS), and goat polyclonal anti-stearoyl CoA desaturase (SCD) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse monoclonal anti-HCV NS5A antibody was obtained from Virogen (Watertown, MA, USA). Polyclonal antibodies specific to mTOR, lipin-1, and acetyl CoA carboxylase (ACACA) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Scrambled miRNA, miR-99a-5p mimics, and miR-99a-5p inhibitor were synthesized and obtained from Genolution Pharmaceuticals (Seoul, Korea). We used miR-99a-5p because miR-99a-5p is a dominant arm among the two strands of miRNA duplex. Scrambled small interfering RNA (siRNA) and siRNA targeting mTOR were obtained from Cell Signaling Technology, Inc. Cells were transfected with miRNA mimics or siRNAs using lipofectamine^TM 2000 (Thermo-Fisher Scientific, MA, USA), according to the manufacturer’s instructions at a final concentration of 50 nM. pcDNA3_flag_mTOR and pcDNA3.1_flag_SREBP-1c were purchased from Addgene (Cambridge, MA, USA).