Ab182561

At PN7.5,4 days after tamoxifen induction, the apical thirds of first mandibular molars from Gli1-CreER;Arid1a^fl/fl mice and littermate controls were cut into pieces and homogenized in RIPA buffer (Cell Signaling, 9806s) supplemented with protease inhibitor (Thermo Fisher Scientific, A32959). Western blot was performed per standard protocol and signals were detected using Azure 300 (Azure biosystems). The primary antibodies are listed in Table S1. HRP-conjugated secondary antibodies (R&D, HAF007, HAF008, and HAF016) were used in the study. For co-immunoprecipitation (co-IP), DPCs cultured in vitro were harvested and lysed in lysis buffer [50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 2 mM EDTA (pH 8.0), 1mM PMSF, 1% NP-40, 5% glycerol]. Then lysates were subjected to immunoprecipitation with anti-Arid1a antibody (Abcam, ab182561) or normal Rabbit IgG (Cell Signaling Technology, 2729) and protein A-Sepharose (VWR, CA97067-898). Immune complexes were washed and subjected to immunoblotting with anti-Arid1a (Santa Cruz, sc-32761) or anti-Plagl1 (Santa Cruz, sc-166944) antibodies.