anti human mouse brd4 Search Results


gene exp brd4 hs04188087 m1  (Thermo Fisher)
85
Thermo Fisher gene exp brd4 hs04188087 m1
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Gene Exp Brd4 Hs04188087 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc05280004-486-72--1?v=Thermo+Fisher
Average 85 stars, based on 1 article reviews
gene exp brd4 hs04188087 m1 - by Bioz Stars, 2026-08
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human histone h3 sino biological 100005 mm01 β actin mouse monoclonal antibody sungene biotech km9001t anti brd4 antibody santa cruz sc  (Sino Biological)
94
Sino Biological human histone h3 sino biological 100005 mm01 β actin mouse monoclonal antibody sungene biotech km9001t anti brd4 antibody santa cruz sc
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Human Histone H3 Sino Biological 100005 Mm01 β Actin Mouse Monoclonal Antibody Sungene Biotech Km9001t Anti Brd4 Antibody Santa Cruz Sc, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc09546576__pnas__2204716119__sapp-238-66-69?v=Sino+Biological
Average 94 stars, based on 1 article reviews
human histone h3 sino biological 100005 mm01 β actin mouse monoclonal antibody sungene biotech km9001t anti brd4 antibody santa cruz sc - by Bioz Stars, 2026-08
94/100 stars
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brd4  (Bethyl)
96
Bethyl brd4
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Brd4, supplied by Bethyl, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/ppr0573229-360-35-43?v=Bethyl
Average 96 stars, based on 1 article reviews
brd4 - by Bioz Stars, 2026-08
96/100 stars
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mouse monoclonal anti brd4  (Proteintech)
94
Proteintech mouse monoclonal anti brd4
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Mouse Monoclonal Anti Brd4, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc12106687-244-191-199?v=Proteintech
Average 94 stars, based on 1 article reviews
mouse monoclonal anti brd4 - by Bioz Stars, 2026-08
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anti human p brd4  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology anti human p brd4
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Anti Human P Brd4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc07156724__41467_2020_15290_MOESM1_ESM-288-18-32?v=Santa+Cruz+Biotechnology
Average 93 stars, based on 1 article reviews
anti human p brd4 - by Bioz Stars, 2026-08
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bl 149 2h5  (Bethyl)
95
Bethyl bl 149 2h5
<t>BRD4</t> disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.
Bl 149 2h5, supplied by Bethyl, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc09977177-241-28-29?v=Bethyl
Average 95 stars, based on 1 article reviews
bl 149 2h5 - by Bioz Stars, 2026-08
95/100 stars
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anti brd4  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti brd4
a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for <t>BRD4:</t> ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.
Anti Brd4, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc09605963-471-7-32?v=Cell+Signaling+Technology+Inc
Average 96 stars, based on 1 article reviews
anti brd4 - by Bioz Stars, 2026-08
96/100 stars
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virus strains brd4 expression vector  (Addgene inc)
93
Addgene inc virus strains brd4 expression vector
a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for <t>BRD4:</t> ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.
Virus Strains Brd4 Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pm39742810-295-59-65?v=Addgene+inc
Average 93 stars, based on 1 article reviews
virus strains brd4 expression vector - by Bioz Stars, 2026-08
93/100 stars
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rabbit  (Proteintech)
96
Proteintech rabbit
a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for <t>BRD4:</t> ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.
Rabbit, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc11430030-41-21-25?v=Proteintech
Average 96 stars, based on 1 article reviews
rabbit - by Bioz Stars, 2026-08
96/100 stars
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addgene plasmid  (Addgene inc)
93
Addgene inc addgene plasmid
a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for <t>BRD4:</t> ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.
Addgene Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pm39454579-568-71-71?v=Addgene+inc
Average 93 stars, based on 1 article reviews
addgene plasmid - by Bioz Stars, 2026-08
93/100 stars
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addgene rrid addgene 91793 transfected construct  (Addgene inc)
93
Addgene inc addgene rrid addgene 91793 transfected construct
a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for <t>BRD4:</t> ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.
Addgene Rrid Addgene 91793 Transfected Construct, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/10__7554_slash_elife__92764-228-54-55?v=Addgene+inc
Average 93 stars, based on 1 article reviews
addgene rrid addgene 91793 transfected construct - by Bioz Stars, 2026-08
93/100 stars
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anti brd4 rabbit antibody  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc anti brd4 rabbit antibody
Effect of ionizing radiation on the proliferation and apoptosis of ARV-771–treated head and neck squamous cell carcinoma (HNSCC) cells. ( A ) ARV-771–treated SAS and Ca9-22 cells for 24 h were collected and analyzed for bromodomain protein 4 <t>(BRD4)</t> protein expression using Western blotting. ( B – D ) ARV-771–treated cells were irradiated with 6 Gy and cultured for 3 days. Cells were counted, reseeded, and cultured for another 3 days (total, 6 days). The cultured cells for 3 or 6 days were harvested for estimation of proliferation ( B ) and apoptosis analysis ( C , D ). ( B ) Results show the cell proliferation rate calculated from the ratio of the number of viable cells at 3 (for SAS) or 6 days (for Ca9-22) of culture to the number of seeded cells. * p < 0.05. ( C ) Representative cytogram of Annexin V/propidium iodide staining in SAS cells are shown. ( D ) Results are shown as percentage of Annexin V-positive cells. * p < 0.05.
Anti Brd4 Rabbit Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+mouse+brd4/pmc12840072-43-0-44?v=Cell+Signaling+Technology+Inc
Average 94 stars, based on 1 article reviews
anti brd4 rabbit antibody - by Bioz Stars, 2026-08
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Image Search Results


BRD4 disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Inhibition of Bromodomain and Extraterminal Domain Family Proteins Ameliorates Experimental Renal Damage

doi: 10.1681/ASN.2015080910

Figure Lengend Snippet: BRD4 disruption inhibits TNF-α–induced upregulation of proinflammatory genes in renal tubular epithelial cells. HK2 cells were transfected with a pool of BRD4 siRNA or nonspecific scramble siRNA (40 nM) for 48 hours. Cells were then treated with TNF-α (5 ng/ml) for 3 hours more. Gene expression of BRD4 and the proinflammatory genes CCL-2, CCL-5 and IL-6 was measured by real–time quantitative PCR. Data are representative of four independent experiments, and results are expressed as the means±SEM. *P<0.05 versus control untransfected cells; #P<0.05 versus nonspecific scramble siRNA.

Article Snippet: The mRNA copy numbers were calculated for each sample by the instrument software using the Ct value (arithmetic fit point analysis for the lightcycler). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Mouse or Human Assay Identification Mouse assay identification (FAM) Ccl-2 Mm00441242_m1 Ccl-5 Mm01302428_m1 Il-6 Mm00446190_m1 Ngal Mm01324470_m1 Kim-1 Mm00506686_m1 Mouse assay identification (VIC) gapdh Mm99999915_g1 Human assay identification (FAM) CCL-2 Hs00234140_m1 CCL-5 Hs00174575_m1 IL-6 Hs00174131_m1 BRD2 Hs01121986_g1 BRD4 Hs04188087_m1 LTB Hs00242739_m1 CSF2 Hs00929873_m1 CCL-20 Hs01011368_m1 NFKB1A ( I κ B α) Hs00355671_g1 CXCL-16 Hs00222859_m1 Human assay identification (VIC) 18s eukaryotic ribosomal 4210893E GAPDH Hs02758991_g1 Open in a separate window Gene assays

Techniques: Transfection, Expressing, Real-time Polymerase Chain Reaction

JQ1 inhibits the direct binding of BRD4 at the CCL-2, CCL-5, and IL-6 promoters through histone acetylation in renal tubular epithelial cells. HK2 cells were pretreated with (−)JQ1 or JQ1 (500 nM) for 1 hour and further stimulated with TNF-α (5 ng/ml) for 3 hours more. ChIP assays were performed using specific antibodies against (A) BRD4, (B) AcH3, and (C) AcH4. Normal rabbit IgG was used as a negative control. Enrichment of BRD4–specific DNA sequences around the transcriptional sites of the human CCL-2, CCL-5, and IL-6 promoters was measured by real–time quantitative PCR using specific primers. Data from three independent experiments are shown, in which each quantitative PCR was run in triplicate. Results are represented as the relative enrichment of BRD4 binding compared with the negative control antibody (IgG). *P<0.05 versus corresponding IgG; #P<0.05 versus (−)JQ1-treated cells.

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Inhibition of Bromodomain and Extraterminal Domain Family Proteins Ameliorates Experimental Renal Damage

doi: 10.1681/ASN.2015080910

Figure Lengend Snippet: JQ1 inhibits the direct binding of BRD4 at the CCL-2, CCL-5, and IL-6 promoters through histone acetylation in renal tubular epithelial cells. HK2 cells were pretreated with (−)JQ1 or JQ1 (500 nM) for 1 hour and further stimulated with TNF-α (5 ng/ml) for 3 hours more. ChIP assays were performed using specific antibodies against (A) BRD4, (B) AcH3, and (C) AcH4. Normal rabbit IgG was used as a negative control. Enrichment of BRD4–specific DNA sequences around the transcriptional sites of the human CCL-2, CCL-5, and IL-6 promoters was measured by real–time quantitative PCR using specific primers. Data from three independent experiments are shown, in which each quantitative PCR was run in triplicate. Results are represented as the relative enrichment of BRD4 binding compared with the negative control antibody (IgG). *P<0.05 versus corresponding IgG; #P<0.05 versus (−)JQ1-treated cells.

Article Snippet: The mRNA copy numbers were calculated for each sample by the instrument software using the Ct value (arithmetic fit point analysis for the lightcycler). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Mouse or Human Assay Identification Mouse assay identification (FAM) Ccl-2 Mm00441242_m1 Ccl-5 Mm01302428_m1 Il-6 Mm00446190_m1 Ngal Mm01324470_m1 Kim-1 Mm00506686_m1 Mouse assay identification (VIC) gapdh Mm99999915_g1 Human assay identification (FAM) CCL-2 Hs00234140_m1 CCL-5 Hs00174575_m1 IL-6 Hs00174131_m1 BRD2 Hs01121986_g1 BRD4 Hs04188087_m1 LTB Hs00242739_m1 CSF2 Hs00929873_m1 CCL-20 Hs01011368_m1 NFKB1A ( I κ B α) Hs00355671_g1 CXCL-16 Hs00222859_m1 Human assay identification (VIC) 18s eukaryotic ribosomal 4210893E GAPDH Hs02758991_g1 Open in a separate window Gene assays

Techniques: Binding Assay, Negative Control, Real-time Polymerase Chain Reaction

JQ1 attenuates gene expression of proinflammatory factors in obstructed kidneys. Mice were treated with JQ1 (100 or 60 mg/kg per day) or vehicle (10% cyclodextran solution) starting 24 hours before UUO and studied after 2 or 5 days. (A and B) RNA was isolated from frozen samples of whole kidney, and gene expression levels were evaluated by real–time quantitative PCR. (C) CCL-2 protein levels were evaluated by ELISA. Results are means±SEM of six to eight animals per group. Data are normalized versus contralateral untreated kidney (considered as one). (D) BRD4 downregulates the proinflammatory gene expression in the damaged kidney. (E) A ChIP assay was carried out in renal samples from UUO mice treated or not treated with JQ1 using an antibody specific for BRD4 or normal rabbit IgG, the latter being a negative control. Enrichment of BRD4 binding regions in the promoters of mouse Ccl-2, Ccl-5, and Il-6 genes was quantified by quantitative PCR using specific primers. Data are from two independent experiments, and each quantitative PCR was run in triplicate. Results are expressed as the n-fold enrichment of anti-BRD4 antibody relative to a negative control antibody (normal rabbit IgG) and further normalized versus contralateral kidney (considered as one). *P<0.05 versus contralateral (C); #P<0.05 versus vehicle–treated obstructed (Ob) kidneys.

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Inhibition of Bromodomain and Extraterminal Domain Family Proteins Ameliorates Experimental Renal Damage

doi: 10.1681/ASN.2015080910

Figure Lengend Snippet: JQ1 attenuates gene expression of proinflammatory factors in obstructed kidneys. Mice were treated with JQ1 (100 or 60 mg/kg per day) or vehicle (10% cyclodextran solution) starting 24 hours before UUO and studied after 2 or 5 days. (A and B) RNA was isolated from frozen samples of whole kidney, and gene expression levels were evaluated by real–time quantitative PCR. (C) CCL-2 protein levels were evaluated by ELISA. Results are means±SEM of six to eight animals per group. Data are normalized versus contralateral untreated kidney (considered as one). (D) BRD4 downregulates the proinflammatory gene expression in the damaged kidney. (E) A ChIP assay was carried out in renal samples from UUO mice treated or not treated with JQ1 using an antibody specific for BRD4 or normal rabbit IgG, the latter being a negative control. Enrichment of BRD4 binding regions in the promoters of mouse Ccl-2, Ccl-5, and Il-6 genes was quantified by quantitative PCR using specific primers. Data are from two independent experiments, and each quantitative PCR was run in triplicate. Results are expressed as the n-fold enrichment of anti-BRD4 antibody relative to a negative control antibody (normal rabbit IgG) and further normalized versus contralateral kidney (considered as one). *P<0.05 versus contralateral (C); #P<0.05 versus vehicle–treated obstructed (Ob) kidneys.

Article Snippet: The mRNA copy numbers were calculated for each sample by the instrument software using the Ct value (arithmetic fit point analysis for the lightcycler). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Mouse or Human Assay Identification Mouse assay identification (FAM) Ccl-2 Mm00441242_m1 Ccl-5 Mm01302428_m1 Il-6 Mm00446190_m1 Ngal Mm01324470_m1 Kim-1 Mm00506686_m1 Mouse assay identification (VIC) gapdh Mm99999915_g1 Human assay identification (FAM) CCL-2 Hs00234140_m1 CCL-5 Hs00174575_m1 IL-6 Hs00174131_m1 BRD2 Hs01121986_g1 BRD4 Hs04188087_m1 LTB Hs00242739_m1 CSF2 Hs00929873_m1 CCL-20 Hs01011368_m1 NFKB1A ( I κ B α) Hs00355671_g1 CXCL-16 Hs00222859_m1 Human assay identification (VIC) 18s eukaryotic ribosomal 4210893E GAPDH Hs02758991_g1 Open in a separate window Gene assays

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Negative Control, Binding Assay

Gene assays

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Inhibition of Bromodomain and Extraterminal Domain Family Proteins Ameliorates Experimental Renal Damage

doi: 10.1681/ASN.2015080910

Figure Lengend Snippet: Gene assays

Article Snippet: The mRNA copy numbers were calculated for each sample by the instrument software using the Ct value (arithmetic fit point analysis for the lightcycler). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Mouse or Human Assay Identification Mouse assay identification (FAM) Ccl-2 Mm00441242_m1 Ccl-5 Mm01302428_m1 Il-6 Mm00446190_m1 Ngal Mm01324470_m1 Kim-1 Mm00506686_m1 Mouse assay identification (VIC) gapdh Mm99999915_g1 Human assay identification (FAM) CCL-2 Hs00234140_m1 CCL-5 Hs00174575_m1 IL-6 Hs00174131_m1 BRD2 Hs01121986_g1 BRD4 Hs04188087_m1 LTB Hs00242739_m1 CSF2 Hs00929873_m1 CCL-20 Hs01011368_m1 NFKB1A ( I κ B α) Hs00355671_g1 CXCL-16 Hs00222859_m1 Human assay identification (VIC) 18s eukaryotic ribosomal 4210893E GAPDH Hs02758991_g1 Open in a separate window Gene assays

Techniques: Mouse Assay

a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for BRD4: ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: METTL3 acetylation impedes cancer metastasis via fine-tuning its nuclear and cytosolic functions

doi: 10.1038/s41467-022-34209-5

Figure Lengend Snippet: a Polysome fractionation analysis in METTL3 reconstituting MDA-MB-231 cells. Absorbance was continuously measured at 260 nm. Representative profile of two independently performed experiments with similar results. b SUnSET assays show enhanced protein production in METTL3 K177Q reconstituting MDA-MB-231 cells. Cell lysates were extracted and probed the indicated antibodies. c Scatter plot of translation efficiency. The read number from the indicated reconstituting cells was calculated by the ratio of the polysome fraction to the read number in the sub-polysome fraction (40/60/80 S). d Scatter plot of RNA-seq data. The average read number from METTL3 WT versus METTL3 K177Q MDA-MB-231 cells (left panel), and average read number from CTRL versus METTL3-deficient MDA-MB-231 cells (right panel) are shown. e Relative levels of the indicated mRNAs in METTL3 reconstituting MDA-MB-231 cells measured by QRT-PCR analysis ( n = 3 biologically independent experiments). From left to right for PTPN14: ns P = 0.084, ns P = 0.45, ns P = 0.75, respectively, by two-sided t test. From left to right for LPP: *** P = 0.0004, ns P = 0.24, ns P = 0.38, respectively, by two-sided t test. From left to right for BRD4: ns P = 0.81, ns P = 0.06, ns P = 0.80, respectively, by two-sided t test. f QRT-PCR quantification of the indicated mRNAs in polysome fractions presented as the ratio to the relative mRNA levels in sub-polysome fractions ( n = 3 biologically independent experiments). From left to right: ** P = 0.0099, ** P = 0.0047, * P = 0.012, respectively, by two-sided t test. g QRT-PCR quantification of METTL3-associated mRNAs in MDA-MB-231 cells ( n = 3 biologically independent experiments). From left to right: * P = 0.015, * P = 0.037, ** P = 0.0022, respectively, by two-sided t test. h Lysates of METTL3 reconstituting MDA-MB-231 cells infected with the indicated lentiviruses were subjected to IB analysis. i Relative mRNA levels of the indicated mRNAs in MDA-MB-231 cells infected with the indicated lentiviruses were analyzed by QRT-PCR ( n = 3 biologically independent experiments). From left to right: ns P = 0.68, ns P = 0.53, ns P = 0.21, respectively, by two-sided t test. j Lysates of METTL3 reconstituting MDA-MB-231 cells were treated with RNase A, followed by IP and IB analysis. All data are represented as mean ± SD. All P values were calculated by Student’s t test. Source data are provided as a Source Data file.

Article Snippet: Anti-acetylated Lys (9441, 1:1000), anti-p300 (86377, 1:1000), anti-BRD4 (13440, 1:1000), anti-Myc-tag (mouse, 2276, Clone No. 9B11, 1:10000 for WB; 1:800 for IF), anti-AMPKα (2532, 1:1000) and anti-phospho-AMPKα-T172 (2535, 1:1000) were obtained from Cell Signaling Technology.

Techniques: Fractionation, RNA Sequencing Assay, Quantitative RT-PCR, Infection

Effect of ionizing radiation on the proliferation and apoptosis of ARV-771–treated head and neck squamous cell carcinoma (HNSCC) cells. ( A ) ARV-771–treated SAS and Ca9-22 cells for 24 h were collected and analyzed for bromodomain protein 4 (BRD4) protein expression using Western blotting. ( B – D ) ARV-771–treated cells were irradiated with 6 Gy and cultured for 3 days. Cells were counted, reseeded, and cultured for another 3 days (total, 6 days). The cultured cells for 3 or 6 days were harvested for estimation of proliferation ( B ) and apoptosis analysis ( C , D ). ( B ) Results show the cell proliferation rate calculated from the ratio of the number of viable cells at 3 (for SAS) or 6 days (for Ca9-22) of culture to the number of seeded cells. * p < 0.05. ( C ) Representative cytogram of Annexin V/propidium iodide staining in SAS cells are shown. ( D ) Results are shown as percentage of Annexin V-positive cells. * p < 0.05.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: Effect of ionizing radiation on the proliferation and apoptosis of ARV-771–treated head and neck squamous cell carcinoma (HNSCC) cells. ( A ) ARV-771–treated SAS and Ca9-22 cells for 24 h were collected and analyzed for bromodomain protein 4 (BRD4) protein expression using Western blotting. ( B – D ) ARV-771–treated cells were irradiated with 6 Gy and cultured for 3 days. Cells were counted, reseeded, and cultured for another 3 days (total, 6 days). The cultured cells for 3 or 6 days were harvested for estimation of proliferation ( B ) and apoptosis analysis ( C , D ). ( B ) Results show the cell proliferation rate calculated from the ratio of the number of viable cells at 3 (for SAS) or 6 days (for Ca9-22) of culture to the number of seeded cells. * p < 0.05. ( C ) Representative cytogram of Annexin V/propidium iodide staining in SAS cells are shown. ( D ) Results are shown as percentage of Annexin V-positive cells. * p < 0.05.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Expressing, Western Blot, Irradiation, Cell Culture, Staining

Effects of ARV-771 or BRD4 knockdown on radiosensitivity of HNSCC cells. ( A ) ARV-771–treated HNSCC cells were irradiated with X-rays, and cells were collected at 24 h following irradiation for colony assay. Results show survival rates at each dose with respect to non-irradiated cells. * p < 0.05. ( B ) SAS and Ca9-22 cells transfected with siRNA targeting BRD4 were harvested for BRD4 protein expression analysis by Western blotting. ( C ) BRD4-knockdown HNSCC cells were irradiated with X-rays, and cells were collected at 24 h following irradiation for colony assay. Results show survival rates at each dose with respect to non-irradiated cells. * p < 0.05.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: Effects of ARV-771 or BRD4 knockdown on radiosensitivity of HNSCC cells. ( A ) ARV-771–treated HNSCC cells were irradiated with X-rays, and cells were collected at 24 h following irradiation for colony assay. Results show survival rates at each dose with respect to non-irradiated cells. * p < 0.05. ( B ) SAS and Ca9-22 cells transfected with siRNA targeting BRD4 were harvested for BRD4 protein expression analysis by Western blotting. ( C ) BRD4-knockdown HNSCC cells were irradiated with X-rays, and cells were collected at 24 h following irradiation for colony assay. Results show survival rates at each dose with respect to non-irradiated cells. * p < 0.05.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Knockdown, Irradiation, Colony Assay, Transfection, Expressing, Western Blot

γH2AX and BRD4 localization and effects of ARV-771 or BRD4 knockdown γH2AX expression in irradiated SAS cells. ( A , B ) SAS cells were irradiated with 2 Gy and cultured for 30 min. Samples are analyzed using immunofluorescence staining. ( A ) DAPI (nuclei) and γH2AX in non-irradiated and 2 Gy–irradiated SAS cells. ( B ) Expression of BRD4 (green) and γH2AX (red) in 2 Gy–irradiated SAS cells. The white dotted line in the figure indicates the outline of the nucleus. ( C , D ) ARV-771–treated ( C ) or BRD4 knockdown ( D ) SAS cells were irradiated with X-rays, and cells were collected at 15 min, 30 min, and 3 h following irradiation for an analysis of γH2AX expression by Western blotting.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: γH2AX and BRD4 localization and effects of ARV-771 or BRD4 knockdown γH2AX expression in irradiated SAS cells. ( A , B ) SAS cells were irradiated with 2 Gy and cultured for 30 min. Samples are analyzed using immunofluorescence staining. ( A ) DAPI (nuclei) and γH2AX in non-irradiated and 2 Gy–irradiated SAS cells. ( B ) Expression of BRD4 (green) and γH2AX (red) in 2 Gy–irradiated SAS cells. The white dotted line in the figure indicates the outline of the nucleus. ( C , D ) ARV-771–treated ( C ) or BRD4 knockdown ( D ) SAS cells were irradiated with X-rays, and cells were collected at 15 min, 30 min, and 3 h following irradiation for an analysis of γH2AX expression by Western blotting.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Knockdown, Expressing, Irradiation, Cell Culture, Immunofluorescence, Staining, Western Blot

γH2AX and BRD4 or ΔNp63 localization in irradiated SAS cells. SAS cells were irradiated with 2 Gy and cultured for 30 min. Samples were analyzed using immunofluorescence staining. ( Left panel ) Expression of BRD4 (green) and γH2AX (red) in 2 Gy–irradiated cells. ( Right panel ) Expression of ΔNp63 (green) and γH2AX (red) in 2 Gy–irradiated cells.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: γH2AX and BRD4 or ΔNp63 localization in irradiated SAS cells. SAS cells were irradiated with 2 Gy and cultured for 30 min. Samples were analyzed using immunofluorescence staining. ( Left panel ) Expression of BRD4 (green) and γH2AX (red) in 2 Gy–irradiated cells. ( Right panel ) Expression of ΔNp63 (green) and γH2AX (red) in 2 Gy–irradiated cells.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Irradiation, Cell Culture, Immunofluorescence, Staining, Expressing

Protein expression of BRD4 and ΔNp63α treated with ΔNp63 knockdown, ARV-771, and BRD4 knockdown cells. ( A ) ΔNp63 knockdown, ( B ) ARV-771–treated, ( C ) BRD4 knockdown SAS cells were harvested for Western blotting to analyze BRD4 and ΔNp63α protein expression.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: Protein expression of BRD4 and ΔNp63α treated with ΔNp63 knockdown, ARV-771, and BRD4 knockdown cells. ( A ) ΔNp63 knockdown, ( B ) ARV-771–treated, ( C ) BRD4 knockdown SAS cells were harvested for Western blotting to analyze BRD4 and ΔNp63α protein expression.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Expressing, Knockdown, Western Blot

A proposed working model of BRD4-dependent regulation of DNA damage responses. Under a condition that BRD4 intact, BRD4-associated SEs support transcriptional programs involved in cell survival and DNA repair, contributing to efficient DNA damage repair and relative radioresistance. ARV-771 treatment induces pharmacological degradation of BRD4 and disorganization of SEs, leading to enhanced γH2AX signaling, transcriptional stress, and increased radiosensitization. In contrast, BRD4 knockdown alters SE function without inducing acute SE degradation, resulting in reduced γH2AX levels following irradiation and a distinct mode of radiosensitization. This model proposes that γH2AX reflects different biological processes depending on the mode of BRD4 suppression.

Journal: Current Issues in Molecular Biology

Article Title: Role of the Super-Enhancer Component Bromodomain Protein 4 in the Radiation Response of Human Head and Neck Squamous Cell Carcinoma Cells

doi: 10.3390/cimb48010071

Figure Lengend Snippet: A proposed working model of BRD4-dependent regulation of DNA damage responses. Under a condition that BRD4 intact, BRD4-associated SEs support transcriptional programs involved in cell survival and DNA repair, contributing to efficient DNA damage repair and relative radioresistance. ARV-771 treatment induces pharmacological degradation of BRD4 and disorganization of SEs, leading to enhanced γH2AX signaling, transcriptional stress, and increased radiosensitization. In contrast, BRD4 knockdown alters SE function without inducing acute SE degradation, resulting in reduced γH2AX levels following irradiation and a distinct mode of radiosensitization. This model proposes that γH2AX reflects different biological processes depending on the mode of BRD4 suppression.

Article Snippet: Anti-BRD4 rabbit antibody (#63759), anti-deltaN p63 (E6Q3O) rabbit antibody (##67825), anti-Glyceraldehyde–3–phosphate dehydrogenase (GAPDH) rabbit antibody (#5174), anti-p-histone H2A.X (γH2AX; #9718), horseradish peroxidase-conjugated anti-rabbit IgG antibody (#7074), horseradish peroxidase-conjugated anti-mouse IgG antibody (#7076), and AlexaFluor 488 ® -conjugated mouse IgG antibody (#4408) were purchased from Cell Signaling Technology Japan, K.K. (Tokyo, Japan).

Techniques: Knockdown, Irradiation