Anti bcl6 sc 858

Mice were euthanized on day 15 following X-31 infection. Mediastinal lymph nodes were removed and fixed in 4% formaldehyde, embedded in paraffin, sectioned, and stained with anti-Bcl6 (sc-858; Santa Cruz) antibodies. Images were acquired with a Nikon TiE microscope equipped with a 10×, 0.3 NA objective, motorized stage, and DS-Ri2 CMOS camera. Image capture, processing, and analysis were performed using NIS Elements software (Nikon Instruments).

Serum levels of HBsAg and HBeAg in the DNA-injected mice were determined by HBsAg II kit (Cat:04687787190, Roche, Switzerland) and HBeAg kit (11820583122, Roche), respectively and measured by an automated analyzer (Roche Cobas 6000). DNAs transfection in the cells was performed by Lipofectamine^TM 3000 (Invitrogen) according to the manufacturer's instructions. For western blot analysis, cells were lysed with RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% NP40, 0.1% SDS, 0.1% deoxycholate and 2 mM EDTA) containing protease inhibitors (5 μg/ml aprotinin, 10 μg/ml leupeptin, 1 mM PMSF) and the antibodies used included anti-HBcAg (LTK BioLaboratories, Taiwan), anti-HBsAg (A10F1, kindly provided by Dr. Sheng-Chung Lee, National Taiwan University, Taiwan), anti-BCL6 (sc-858, Santa Cruz, USA), anti-β-actin (A5441, Sigma, USA) and horse radish peroxidase (HRP)-conjugated sheep anti-rabbit, or anti-mouse IgG (GE healthcare, USA). The cytokines used for stimulating Bcl6 expression or chemokine induction included TNF-α (315-01A, PeProtech, USA) and IFN-γ (485MI, R&D, USA). The luciferase activity was measured by Dual-Glo® Luciferase Assay System (Promega, USA) following the manufacturer's instructions.