Anti hcv core c7 50

Proteins extracted from cells or concentrated from elutriants were analyzed with SDS-PAGE, the proteins on the membrane were probed with anti-HCV Core (C7-50, Abcam Ltd.), anti-NS3 (H23, Abcam Ltd.), anti-hA3G (ab75560, Abcam Ltd.), anti-HA (6E2, Cell Signaling Biotechnology Inc.), or anti-HA-tag [HRP] (A00169, GenScript.) antibody, respectively, with anti-Actin antibody (TA-09, ZSGB-BIO, China) served as the control. After washing with TBST, the membrane was incubated with goat anti-mouse (sc-2005, Santa Cruz Biotechnology Inc.) or goat anti-rabbit (sc-2004, Santa Cruz Biotechnology Inc.) secondary antibody, respectively. Protein signals were visualized and captured using Immobilon Western Chemiluminescent HRP Substrate ECL working solution (Millipore Inc.) with ChemiDo XRS gel imager system (Bio-Rad, CA).

HCV core 4a and core 4f genes were amplified by PCR from pCI/core-4aR, pCI/core-4fC plasmid (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6060129/) (accessed on 12 April 2022) and placed downstream of a transthyretin (TTR) liver-specific promoter [32 (link)]. Core 4a and core 4f genes was inserted into the StuI site of the pTTR1-ExV3 plasmid, and the transgenes were prepared by purifying the HindIII fragment containing TTR promoter and the core 4a/core 4f coding sequence. The TTR promoter was kindly provided by Dr. Iannis Talianidis, Institute of Molecular Biology and Biotechnology of FORTH in Crete, Heraklion, Greece. Transgenic CBA-C57BL/6 mice, which express liver-specifically core 4a (TTRcore4a) and core 4f (TTRcore4f) proteins, were generated in the Transgenesis Facility of the Biomedical Sciences Research Center “Alexander Fleming”, Vari, Greece. Mice were maintained under specific pathogen-free (SPF) conditions. Western blot using anti-GAPDH (6C5, Abcam, Cambridge, MA, USA) and anti-HCV core (C7-50, Abcam, Cambridge, MA, USA) antibodies was conducted according to the manufacturer’s general guidelines.