Pxr antibody

Dulbecco’s Modified Eagle Medium (DMEM) (Gibco), fetal bovine serum (Gibco), penicillin-streptomycin (pen/strep) (Gibco), trypsin-EDTA (Gibco), histone antibodies (Cell Signalling Technology), GAPDH antibody (Abgenex), TIP60 antibody (Santa Cruz Biotechnology), PXR antibody (Santa Cruz Biotechnology), β-actin antibody (Santa Cruz Biotechnology), lipofectamine 2000 (Invitrogen), lipofectamine RNAiMAX reagent (Invitrogen), Wizard SV gel and PCR clean up system (Promega), Ni-nitrilotriacetic acid (Ni-NTA) (Genetix), histone 4 peptide (Millipore), Alexa Fluor 594 phalloidin (Invitrogen), NU9056 (Tocris), Verso cDNA synthesis kit (Thermo Scientific), SYBR green (Applied Biosystems), TRIzol reagent (Life technologies), DAPI antifade mountant (Invitrogen), horseradish peroxidase–conjugated heavy chain secondary mice and rabbit antibodies (Santa Cruz Biotechnology), light chain–specific horseradish peroxidase–conjugated secondary mice antibody (Merck), and Protein A-Sepharose 4B conjugate bead (Invitrogen). Primers used in the study were synthesized from Integrated DNA technologies and Eurofins Genomics. Primer sequences are mentioned in supporting information (Table S2). siRNA for TIP60 was purchased from Santa Cruz Biotechnology, while siGL2 (used as control) was synthesized from Sigma, with target sequence ‘CGTACGCGGAATACTTCGA’ (24 (link)).

For Western blotting, transfected cells were harvested and lysed in lysis buffer (50 mM Tris-Cl (pH-8.0), 150 mM NaCl, 1 mM DTT, 5 mM EDTA, 0.5% NP-40 and 1X protease inhibitor cocktail (Amresco) followed by sonication. Protein samples were resolved by SDS-PAGE, transferred onto the methanol-charged PVDF membrane using semi-dry transfer apparatus. Subsequently, the blots were probed with indicated primary antibodies. RFP antibody, GFP antibody, GST antibody, PXR antibody, His antibody and acetylated-lysine antibody were purchased from Santa Cruz Biotechnologies. TIP60 antibody, RXRα antibody and β-actin antibody were procured from Cell Signaling Technology (CST). Acetylated Histone H4 and Histone antibodies were purchased from Millipore. Horseradish peroxidase-conjugated (HRP) anti-rabbit and anti-mouse secondary antibodies were purchased from Santa Cruz Biotechnologies while light chain specific HRP-conjugated anti-rabbit and anti-mouse secondary antibodies were purchased from Millipore. Blots were developed using ECL reagent (BioRad) and C digit scanner (Licor).