rabbit polyclonal anti pan brd4  (Millipore)

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Recruitment of Brd3 and Brd4 to acetylated chromatin is essential for proinflammatory cytokine-induced matrix-degrading enzyme expression

doi: 10.1186/s13018-019-1091-3

Figure Lengend Snippet: The protein expression of the BET family members and the effect of siRNAs in SW1353 cells. a The protein expression of Brd2, Brd3, and Brd4 was confirmed by Western blot assay in SW1353 cells and primary human chondrocytes, respectively. PHC, primary human chondrocyte. b – d SW1353 cells were treated with scrambled siRNA or specific siRNA targeting BRD2 , BRD3 , and BRD4 for 48 h and then treated with or without IL-1β (10 ng/ml) or TNF-α (10 ng/ml) for 6 h. The presence of Brd2, Brd3, and Brd4 proteins in SW1353 cells was confirmed by Western blot assay

Article Snippet: The primary antibodies used were rabbit polyclonal anti-Brd4 (1:1000; Cell Signaling Technology), rabbit monoclonal anti-Brd2 (1:1000; Abcam), mouse monoclonal anti-Brd3 (1:500; Abcam), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000; Santa Cruz Biotechnology, INC).

Techniques: Expressing, Western Blot

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Recruitment of Brd3 and Brd4 to acetylated chromatin is essential for proinflammatory cytokine-induced matrix-degrading enzyme expression

doi: 10.1186/s13018-019-1091-3

Figure Lengend Snippet: The effect of specific siRNAs on regulation of matrix-degrading enzyme gene transcription in SW1353 cells. SW1353 cells were treated with scrambled siRNA or specific siRNA targeting BRD2 , BRD3 , and BRD4 for 48 h and then treated with or without IL-1β (10 ng/ml) or TNF-α (10 ng/ml) for 6 h. a – d The transcriptional expression of MMP1 , MMP3 , MMP13 , and ADAMTS4 was evaluated by real time-PCR. Relative fold-change values were calculated in comparison with the vehicle control that was set to 1 ( n = 3)

Article Snippet: The primary antibodies used were rabbit polyclonal anti-Brd4 (1:1000; Cell Signaling Technology), rabbit monoclonal anti-Brd2 (1:1000; Abcam), mouse monoclonal anti-Brd3 (1:500; Abcam), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000; Santa Cruz Biotechnology, INC).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Recruitment of Brd3 and Brd4 to acetylated chromatin is essential for proinflammatory cytokine-induced matrix-degrading enzyme expression

doi: 10.1186/s13018-019-1091-3

Figure Lengend Snippet: The recruitment of Brd3 and Brd4 to the chromatin responsible for IL-1β- or TNF-α-induced transcription. a – d Human chondrocytes were pretreated with or without I-BET151 (1 μM), followed by addition of vehicle, IL-1β (10 ng/ml) or TNF-α (10 ng/ml) for 6 h, and ChIP assays were performed for Brd3. e – h Human chondrocytes were pretreated with or without I-BET151 (1 μM), followed by addition of vehicle, IL-1β (10 ng/ml) or TNF-α (10 ng/ml) for 6 h, and ChIP assays were performed for Brd4. The quantitative analysis of targeted promoter regions was determined by real-time PCR using specific primers for MMP1 , MMP3 , MMP13 , and ADAMTS4. Relative fold-change values were calculated in comparison with the vehicle control that was set to 1 ( n = 2)

Article Snippet: The primary antibodies used were rabbit polyclonal anti-Brd4 (1:1000; Cell Signaling Technology), rabbit monoclonal anti-Brd2 (1:1000; Abcam), mouse monoclonal anti-Brd3 (1:500; Abcam), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000; Santa Cruz Biotechnology, INC).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Recruitment of Brd3 and Brd4 to acetylated chromatin is essential for proinflammatory cytokine-induced matrix-degrading enzyme expression

doi: 10.1186/s13018-019-1091-3

Figure Lengend Snippet: Schematic mechanism for Brd-mediated transcriptional regulation of MMP1 , MMP3 , MMP13 , and ADAMTS4 genes in human chondrycytes. This model proposes that IL-1β or TNF-α stimulation leads to the H4K5/8/12Ac at TSS regions and the recruitment of Brd3, Brd4, and p-TEFb to these regions. Subsequently, Ser2 RNAP II CTD is phosphorylated by p-TEFb. The recruitment of Brd3 and Brd4 is blocked by I-BET-151

Article Snippet: The primary antibodies used were rabbit polyclonal anti-Brd4 (1:1000; Cell Signaling Technology), rabbit monoclonal anti-Brd2 (1:1000; Abcam), mouse monoclonal anti-Brd3 (1:500; Abcam), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000; Santa Cruz Biotechnology, INC).

Techniques: