Anti gapdh m20006

The rabbit anti-NS2 polyclonal antibody and mouse anti-NP monoclonal antibody were prepared previously locally. Other antibodies were as follows: anti-histone H3.1 (P30266), anti-Flag (m20008) and anti-GAPDH (M20006) from Abmart (Shanghai, China); anti-c-Myc tag (CW0089), FITC-conjugated anti-rabbit IgG (CW0113), cy3-conjugated anti-mouse IgG (CW0159), HRP-conjugated anti-mouse IgG (CW0102) and anti-rabbit IgG (CW0103) from CWBIO (Beijing, China); anti-Crm1 (611832) from BD Transduction Laboratories (Franklin Lakes, NJ, USA); anti-HMGB1 (2600-1), anti-TFIIB (3728-1), anti-tubulin-alpha (2871-1) and anti-CHD3 (2969-1) from EPITOMICS (CA, USA); and normal mouse IgG (sc-2025) from Santa Cruz Biotechnology (CA, USA).

The following antibodies were used for Chromatin Immunoprecipitation (ChIP) or immunoblotting: anti-TCF1(2003S; Cell Signalling Technology), anti-beta-catenin (ab32572; Abcam), anti-Phospho-β-Catenin (9561s; Cell Signalling Technology), anti-GSK3(ab40870; Abcam), anti-GSK3 phospho Y216 + Y279(ab68476; Abcam), anti-H3K4me3 (9751s; Cell Signalling Technology), anti-H3K27ac (8173s; Cell Signalling Technology), anti-H3K27me3 (17–622, Millipore), anti-H3K9me3 (ab8898, Abcam), anti-GAPDH (M20006; Abmart), and anti-Lamin B (66091-1-Ig, Proteintech).
For immunoblotting, cells were lysed for 1 h at 4°C in ice-cold lysis buffer (50 mM HEPES, pH 7.4, 150 mM NaCl, 0.5 mM EGTA, 1% protease inhibitor cocktail [Sigma], 1 mM sodium orthovanadate, 1 mM NaF, 1% [vol/vol] Triton X-100, and 10% [vol/vol] glycerol). After centrifugation for 10 min at 12,000g, the supernatant was boiled in reducing SDS sample loading buffer and subjected by SDS-PAGE. The indicated specific primary antibodies were used, followed by horseradish peroxidase-conjugated goat anti-mouse IgG or goat anti-rabbit IgG (Sigma) as the secondary antibody. Nuclear and cytoplasmic protein fractions were purified using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific) according to the manufacturer’s introduction.