Anti hbc

After transfection, cells were washed with ice-cold PBS after 36 h, harvested by scraping, then lysed using an RIPA lysis buffer (50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 1% NP40, 0.1% SDS) supplemented with a protease inhibitor cocktail (Thermo Scientific). After centrifugation at 13,000 rpm at 4 °C for 5 min, supernatants were incubated with primary antibody or IgG (Santa Cruz, Santa Cruz, CA, USA) and protein G agarose beads (GE Healthcare, Chicago, IL, USA) at 4 °C overnight. Immunoprecipitates were washed with a washing buffer (50 mM Tris-HCl [pH 7.4], 150 mM NaCl) three times, then whole cell lysate and immunoprecipitated fractions were used for western blot analysis. The following primary antibodies were used: anti-HA (catalog no. H6908, Sigma-Aldrich), anti-Flag (catalog no. F1804, Sigma-Aldrich), anti-BAF200 (catalog no. A302-230A, Bethyl), anti-HBc (catalog no. B0586, Dako).

Vendors for antibodies are shown in parentheses: anti-HBc (Dako, Real Carpinteria, CA), anti-STAT3 (GeneTex, Taiwan), anti-GAPDH, anti-tubulin and anti-phospho-Stat3 (tyr705) (Cell Signaling Technology Inc, Danvers, MA). Secondary antibodies include mouse anti-rabbit-HRP, goat anti-mouse-HRP (GeneTex, Taiwan).

Anti-HA-tag (rabbit polyclonal, MilliporeSigma, Burlington, MA, USA), anti-FLAG-tag (mouse monoclonal, MilliporeSigma, Burlington, MA, USA), anti-HA-tag (mouse monoclonal, MilliporeSigma, Burlington, MA, USA), anti-HBc (rabbit polyclonal, DAKO, Carpinteria, CA, USA), and anti-HBc (rabbit monoclonal, Gilead Sciences, Inc., Foster City, CA, USA) were used as primary antibodies. As secondary antibodies conjugated with horseradish peroxidase (HRP), we used goat anti-mouse (MilliporeSigma, Burlington, MA, USA) and goat anti-rabbit (MilliporeSigma, Burlington, MA, USA). For visualization, we used a SuperSignal™ West Femto Maximum Sensitivity Substrate (ThermoFisher Scientific, Waltham, MA, USA) and LAS-4000 imager. As secondary antibodies for the LI-COR system, we used goat anti-rabbit IgG (H + L) IRDye 800CW (LI-COR Biosciences, Lincoln, NE, USA) and goat anti-mouse IgG (H + L) IRDye 680RD (LI-COR Biosciences, Lincoln, NE, USA). Western blots were visualized using an LI-COR Odyssey CLx system and the Image Studio Lite Software (LI-COR Biosciences, Lincoln, NE, USA).