Anti brd4

RAW 264.7 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, U.S.A.). Cell culture medium and Lipofectamine 2000 were from Gibco (Life Technologies, Grand Island, NY, U.S.A.). M-CSF and RANKL recombinant protein factors, FBS, and ELISA kits for TNF-α, IL-1β, and IL-6 were purchased from Thermo Fisher Scientific (Waltham, MA, U.S.A.). I-BET151 was obtained from Santa Cruz Biotechnology (Dallas, Texas, U.S.A.). TRAF6 antibody was purchased from Epitomics, Inc. (Burlingame, CA, U.S.A.) and others including anti-BRD4, anti-p65, anti-IκB-α, anti-NFATc1, anti-OPG, anti-lamin B, and anti-β-actin were all from Cell Signaling Technology (Danvers, MA, U.S.A.). TRACP staining kit was obtained from Nanjing Jiangcheng Bioengineering Institute (Nanjing, China). Non-silencing control siRNA was purchased from QIAGEN (Germantown, MD, U.S.A.) and siRNA targetting BRD4 was synthesized by GenePharma (Shanghai, China). Random hexamer primer and MMLV Reverse Transcriptase were obtained from Promega (Madison, WI, U.S.A.). HotStart SYBR Green qPCR Master Mix (2×) was from TransGen Biotech (Beijing, China). Nitrocellulose membranes were purchased from Millipore (Darmstadt, Germany). Nuclear and Cytoplasmic Protein Extraction Kit was obtained from Beyotime (Jiangsu, China). Other reagents and kits used in the present study were obtained from Sigma (St. Louis, MO, U.S.A.).