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bd2 e49 e460 (MedChemExpress)

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MedChemExpress bd2 e49 e460

a ) Dot plots showing log2 fold enrichment of BRD proteins in the proximal interactome (Turbo-ID) for PRC1 and PRC2 proteins from mouse embryonic stem cells (mESCs), data from . The size of the circle represents the log2 fold enrichment in BRD4 IP relative to IgG control. b ) Like (a) but for enrichment of PRC proteins in BRD4 immunoprecipitation from K562 cells, data from , . The size of the circle represents the t-test difference between the BRD4 IP and the IgG control. c) Immunoblots of endogenous BRD4 IP in H9 hESCs using antibodies that recognise both short and long BRD4 isoforms, with antibodies detecting RING1B, CBX7, CBX4, H3K27ac, H3K23ac, H3K27me3, along with reverse IP with RING1B and MGA antibodies followed by immunoblots for BRD4 and H3K27me3. d ) Immunoblots of GFP-trap co-immunoprecipitation of GFP-BRD4 long isoform (GFP-BRD4L) with Flag-tagged E2F6 and L3MBTL2, HA-tagged EED and EZH2. Immunoblots for β-ACTIN served as controls, e ) Heatmap of CUT&Tag for BRD4, EED, H3K23ac and ChIP-seq data for H3K14ac and RING1B, at active (H3K4me3+), bivalent (H3K4me3+/H3K27me3+) and PRC2 repressed promoters (H3K27me3+). f ) AlphaScreen counts titration of BRD4-BD1 and <t>-BD2</t> interaction with H3K14ac/23ac showing that only BRD4-BD2 interacts with H3K14ac/23ac. Normalized average alpha counts of three replicates were set relative to the highest WT. g) Immunoblots of biotinylated H3K14/K23ac pulldown for N-terminal His-FLAG tagged BRD4 (N-terminal 412 amino acids), in the presence of increasing concentration of iBET-BD2 (iBD2).

Bd2 E49 E460, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 4 article reviews

bd2 e49 e460 - by Bioz Stars, 2026-02

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1) Product Images from "BRD4 represses developmental and neuronal genes through interactions with polycomb complexes"

Article Title: BRD4 represses developmental and neuronal genes through interactions with polycomb complexes

Journal: bioRxiv

doi: 10.64898/2026.01.31.702994

Figure Legend Snippet: a ) Dot plots showing log2 fold enrichment of BRD proteins in the proximal interactome (Turbo-ID) for PRC1 and PRC2 proteins from mouse embryonic stem cells (mESCs), data from . The size of the circle represents the log2 fold enrichment in BRD4 IP relative to IgG control. b ) Like (a) but for enrichment of PRC proteins in BRD4 immunoprecipitation from K562 cells, data from , . The size of the circle represents the t-test difference between the BRD4 IP and the IgG control. c) Immunoblots of endogenous BRD4 IP in H9 hESCs using antibodies that recognise both short and long BRD4 isoforms, with antibodies detecting RING1B, CBX7, CBX4, H3K27ac, H3K23ac, H3K27me3, along with reverse IP with RING1B and MGA antibodies followed by immunoblots for BRD4 and H3K27me3. d ) Immunoblots of GFP-trap co-immunoprecipitation of GFP-BRD4 long isoform (GFP-BRD4L) with Flag-tagged E2F6 and L3MBTL2, HA-tagged EED and EZH2. Immunoblots for β-ACTIN served as controls, e ) Heatmap of CUT&Tag for BRD4, EED, H3K23ac and ChIP-seq data for H3K14ac and RING1B, at active (H3K4me3+), bivalent (H3K4me3+/H3K27me3+) and PRC2 repressed promoters (H3K27me3+). f ) AlphaScreen counts titration of BRD4-BD1 and -BD2 interaction with H3K14ac/23ac showing that only BRD4-BD2 interacts with H3K14ac/23ac. Normalized average alpha counts of three replicates were set relative to the highest WT. g) Immunoblots of biotinylated H3K14/K23ac pulldown for N-terminal His-FLAG tagged BRD4 (N-terminal 412 amino acids), in the presence of increasing concentration of iBET-BD2 (iBD2).

Techniques Used: Control, Immunoprecipitation, Western Blot, ChIP-sequencing, Amplified Luminescent Proximity Homogenous Assay, Titration, Concentration Assay

a ) Heatmap showing BRD4 signal (CPM) for WT and BRD4 BD2 mut1 at protein-coding genes and active enhancers of hESCs. b ) Scatter plot comparing log2 fold change (log2 FC) values for BRD4 BD2-Mut1/WT (X-axis) against BRD4 dTAG/DMSO (Y-axis) conditions. GSEA GO-biological process enrichment lists for genes that are commonly up (red) and down (blue) regulated in both conditions (right). c ) Representative genome browser snapshot displaying signals for RNA-seq WT, BRD4-mutant1, DMSO and dTAGV-1 along with MAX, BRD4, H3K27me3 and H3K4me3. For CUT&Tag (BRD2,3,4, H3K4me3, H3K27me3) and CUT&Run (EED, ser5 Pol-II), the signal is compared as CPM and MAX as ChIP-seq signal from ChIP-atlas. d) Heatmaps displaying H3K27me3 and H3K4me3 ChIP-seq signals along with RNA-seq normalized counts at bivalent genes in WT-H9 and H9-derived BRD4 BD2 mut1 neurons. e ) MA plot illustrating differential gene expression in BRD4 BD2 mut1 compared to WT neurons. Significantly up- and down-regulated bivalent and non-bivalent genes are highlighted in red and blue, respectively. The number of differentially expressed genes with a log2 fold change of 1 and an adjusted p-value of <0.05 is indicated (right). f ) Genome browser tracks showing ChIP-seq data for bivalent histone modifications (H3K4me3 and H3K27me3), fold change over input and RNA-seq (RPKM) for neuronal genes.

Figure Legend Snippet: a ) Heatmap showing BRD4 signal (CPM) for WT and BRD4 BD2 mut1 at protein-coding genes and active enhancers of hESCs. b ) Scatter plot comparing log2 fold change (log2 FC) values for BRD4 BD2-Mut1/WT (X-axis) against BRD4 dTAG/DMSO (Y-axis) conditions. GSEA GO-biological process enrichment lists for genes that are commonly up (red) and down (blue) regulated in both conditions (right). c ) Representative genome browser snapshot displaying signals for RNA-seq WT, BRD4-mutant1, DMSO and dTAGV-1 along with MAX, BRD4, H3K27me3 and H3K4me3. For CUT&Tag (BRD2,3,4, H3K4me3, H3K27me3) and CUT&Run (EED, ser5 Pol-II), the signal is compared as CPM and MAX as ChIP-seq signal from ChIP-atlas. d) Heatmaps displaying H3K27me3 and H3K4me3 ChIP-seq signals along with RNA-seq normalized counts at bivalent genes in WT-H9 and H9-derived BRD4 BD2 mut1 neurons. e ) MA plot illustrating differential gene expression in BRD4 BD2 mut1 compared to WT neurons. Significantly up- and down-regulated bivalent and non-bivalent genes are highlighted in red and blue, respectively. The number of differentially expressed genes with a log2 fold change of 1 and an adjusted p-value of <0.05 is indicated (right). f ) Genome browser tracks showing ChIP-seq data for bivalent histone modifications (H3K4me3 and H3K27me3), fold change over input and RNA-seq (RPKM) for neuronal genes.

Techniques Used: RNA Sequencing, ChIP-sequencing, Derivative Assay, Gene Expression

a) Schematic representation of the protocol used to generate unguided neuronal organoids (UNOs), with images of UNO WT at 5,8, and 41 days. b ) Immunofluorescence images of UNOs at day 41 stained for markers of neuronal progenitor (SOX2), post-mitotic early neurons (TUJ1), scale bars: 100 μm. c ) MA plot for RNA-seq data illustrating differentially expressed genes in day 41 UNOs following 20 hours of BRD4 PROTAC (ZxH) treatment (n=3 independent organoids). d) Geneontology (GO) enrichment analyses of up- and down-regulated genes. e ) Genome browser tracks for normalized reads at TSS for pseudo bulk scCUT&Tag and bulk RNA-seq for immediate early genes (IEGs) upon 20 h BRD4 PROTAC in UNOs (data from (c)). f) UMAP plots stratified by genotype show the annotated cell lineages: WT, BRD4 BD2 mut2, and BRD4 BD2 mut3. Cell clusters are identified by colour, illustrating the contribution of each genotype to specific lineages, such as Glutamatergic, GABAnergic, optic vesicle, and RPE. g) Stacked bar charts for 41-day and 63-day UNOs, detailing the percentage of cells for each annotated cell type across the WT, BRD4 BD2 mut2, and BRD4 BD2 mut3 UNOs. h) Representative bright-field microscopy images of 41-day UNOs, Scale bar=1mm (rest of the images in source file). i) Dot plots showing the average expression level (Z scores) and percentage of cells expressed in Glutamatergic, Diencephalic-1(pink in UMAP), and Diencephalic-2(blue in UMAP), and G2M clusters for bivalent genes that showed significant differential expression in the scRNA-seq data in BRD4-BD2 mut1 and BRD4-BD2 mut2 UNOs.

Figure Legend Snippet: a) Schematic representation of the protocol used to generate unguided neuronal organoids (UNOs), with images of UNO WT at 5,8, and 41 days. b ) Immunofluorescence images of UNOs at day 41 stained for markers of neuronal progenitor (SOX2), post-mitotic early neurons (TUJ1), scale bars: 100 μm. c ) MA plot for RNA-seq data illustrating differentially expressed genes in day 41 UNOs following 20 hours of BRD4 PROTAC (ZxH) treatment (n=3 independent organoids). d) Geneontology (GO) enrichment analyses of up- and down-regulated genes. e ) Genome browser tracks for normalized reads at TSS for pseudo bulk scCUT&Tag and bulk RNA-seq for immediate early genes (IEGs) upon 20 h BRD4 PROTAC in UNOs (data from (c)). f) UMAP plots stratified by genotype show the annotated cell lineages: WT, BRD4 BD2 mut2, and BRD4 BD2 mut3. Cell clusters are identified by colour, illustrating the contribution of each genotype to specific lineages, such as Glutamatergic, GABAnergic, optic vesicle, and RPE. g) Stacked bar charts for 41-day and 63-day UNOs, detailing the percentage of cells for each annotated cell type across the WT, BRD4 BD2 mut2, and BRD4 BD2 mut3 UNOs. h) Representative bright-field microscopy images of 41-day UNOs, Scale bar=1mm (rest of the images in source file). i) Dot plots showing the average expression level (Z scores) and percentage of cells expressed in Glutamatergic, Diencephalic-1(pink in UMAP), and Diencephalic-2(blue in UMAP), and G2M clusters for bivalent genes that showed significant differential expression in the scRNA-seq data in BRD4-BD2 mut1 and BRD4-BD2 mut2 UNOs.

Techniques Used: Immunofluorescence, Staining, RNA Sequencing, Microscopy, Expressing, Quantitative Proteomics

a) UMAP plots show the distribution of single-cell ATAC sequencing (scATAC-seq) data clustered by genotypes WT and BRD4 BD2 mut2 and annotated by cell lineage for WT and BRD4 BD2 mut2. b ) Z-scores (high scores in red and low scores are in blue) showing top transcription factor motifs enriched at Diencephalic, Glutamatergic, G2M and GABAnergic lineages across scATACseq peaks, which are gained in BRD4 BD2 mut 2 UNO compared to WT control. The complete list of enriched TFs is in the source data table.

Figure Legend Snippet: a) UMAP plots show the distribution of single-cell ATAC sequencing (scATAC-seq) data clustered by genotypes WT and BRD4 BD2 mut2 and annotated by cell lineage for WT and BRD4 BD2 mut2. b ) Z-scores (high scores in red and low scores are in blue) showing top transcription factor motifs enriched at Diencephalic, Glutamatergic, G2M and GABAnergic lineages across scATACseq peaks, which are gained in BRD4 BD2 mut 2 UNO compared to WT control. The complete list of enriched TFs is in the source data table.

Techniques Used: Sequencing, Control

Image Search Results

Journal: bioRxiv

Article Title: BRD4 represses developmental and neuronal genes through interactions with polycomb complexes

doi: 10.64898/2026.01.31.702994

Figure Lengend Snippet: a ) Dot plots showing log2 fold enrichment of BRD proteins in the proximal interactome (Turbo-ID) for PRC1 and PRC2 proteins from mouse embryonic stem cells (mESCs), data from . The size of the circle represents the log2 fold enrichment in BRD4 IP relative to IgG control. b ) Like (a) but for enrichment of PRC proteins in BRD4 immunoprecipitation from K562 cells, data from , . The size of the circle represents the t-test difference between the BRD4 IP and the IgG control. c) Immunoblots of endogenous BRD4 IP in H9 hESCs using antibodies that recognise both short and long BRD4 isoforms, with antibodies detecting RING1B, CBX7, CBX4, H3K27ac, H3K23ac, H3K27me3, along with reverse IP with RING1B and MGA antibodies followed by immunoblots for BRD4 and H3K27me3. d ) Immunoblots of GFP-trap co-immunoprecipitation of GFP-BRD4 long isoform (GFP-BRD4L) with Flag-tagged E2F6 and L3MBTL2, HA-tagged EED and EZH2. Immunoblots for β-ACTIN served as controls, e ) Heatmap of CUT&Tag for BRD4, EED, H3K23ac and ChIP-seq data for H3K14ac and RING1B, at active (H3K4me3+), bivalent (H3K4me3+/H3K27me3+) and PRC2 repressed promoters (H3K27me3+). f ) AlphaScreen counts titration of BRD4-BD1 and -BD2 interaction with H3K14ac/23ac showing that only BRD4-BD2 interacts with H3K14ac/23ac. Normalized average alpha counts of three replicates were set relative to the highest WT. g) Immunoblots of biotinylated H3K14/K23ac pulldown for N-terminal His-FLAG tagged BRD4 (N-terminal 412 amino acids), in the presence of increasing concentration of iBET-BD2 (iBD2).

Article Snippet: 1 μg of biotinylated histone H3K14ac/H3K23ac peptide (Cayman Chemicals, Cat. 27520-250ug-CAY) was incubated with 10 μL of streptavidin magnetic beads (Invitrogen 656-01) in 300 μL of binding buffer (50 mM Tris, pH 7.5, 200 mM NaCl and 0.1% NP-40, proteinase inhibitor cocktail) and rotated at room temperature for 30 min. At the same time, FLAG-His tagged BRD4 N -terminal domain containing BD1 and BD2 (E49-E460) (MedChemExpress Cat# HY-P7846), inhibitor of iBET-BD2 (Cayman Chemical Cat# CAY31766), or DMSO were added to the binding buffer on ice.

Techniques: Control, Immunoprecipitation, Western Blot, ChIP-sequencing, Amplified Luminescent Proximity Homogenous Assay, Titration, Concentration Assay

Journal: bioRxiv

Article Title: BRD4 represses developmental and neuronal genes through interactions with polycomb complexes

doi: 10.64898/2026.01.31.702994

Figure Lengend Snippet: a ) Heatmap showing BRD4 signal (CPM) for WT and BRD4 BD2 mut1 at protein-coding genes and active enhancers of hESCs. b ) Scatter plot comparing log2 fold change (log2 FC) values for BRD4 BD2-Mut1/WT (X-axis) against BRD4 dTAG/DMSO (Y-axis) conditions. GSEA GO-biological process enrichment lists for genes that are commonly up (red) and down (blue) regulated in both conditions (right). c ) Representative genome browser snapshot displaying signals for RNA-seq WT, BRD4-mutant1, DMSO and dTAGV-1 along with MAX, BRD4, H3K27me3 and H3K4me3. For CUT&Tag (BRD2,3,4, H3K4me3, H3K27me3) and CUT&Run (EED, ser5 Pol-II), the signal is compared as CPM and MAX as ChIP-seq signal from ChIP-atlas. d) Heatmaps displaying H3K27me3 and H3K4me3 ChIP-seq signals along with RNA-seq normalized counts at bivalent genes in WT-H9 and H9-derived BRD4 BD2 mut1 neurons. e ) MA plot illustrating differential gene expression in BRD4 BD2 mut1 compared to WT neurons. Significantly up- and down-regulated bivalent and non-bivalent genes are highlighted in red and blue, respectively. The number of differentially expressed genes with a log2 fold change of 1 and an adjusted p-value of <0.05 is indicated (right). f ) Genome browser tracks showing ChIP-seq data for bivalent histone modifications (H3K4me3 and H3K27me3), fold change over input and RNA-seq (RPKM) for neuronal genes.

Techniques: RNA Sequencing, ChIP-sequencing, Derivative Assay, Gene Expression

Journal: bioRxiv

Article Title: BRD4 represses developmental and neuronal genes through interactions with polycomb complexes

doi: 10.64898/2026.01.31.702994

Figure Lengend Snippet: a) Schematic representation of the protocol used to generate unguided neuronal organoids (UNOs), with images of UNO WT at 5,8, and 41 days. b ) Immunofluorescence images of UNOs at day 41 stained for markers of neuronal progenitor (SOX2), post-mitotic early neurons (TUJ1), scale bars: 100 μm. c ) MA plot for RNA-seq data illustrating differentially expressed genes in day 41 UNOs following 20 hours of BRD4 PROTAC (ZxH) treatment (n=3 independent organoids). d) Geneontology (GO) enrichment analyses of up- and down-regulated genes. e ) Genome browser tracks for normalized reads at TSS for pseudo bulk scCUT&Tag and bulk RNA-seq for immediate early genes (IEGs) upon 20 h BRD4 PROTAC in UNOs (data from (c)). f) UMAP plots stratified by genotype show the annotated cell lineages: WT, BRD4 BD2 mut2, and BRD4 BD2 mut3. Cell clusters are identified by colour, illustrating the contribution of each genotype to specific lineages, such as Glutamatergic, GABAnergic, optic vesicle, and RPE. g) Stacked bar charts for 41-day and 63-day UNOs, detailing the percentage of cells for each annotated cell type across the WT, BRD4 BD2 mut2, and BRD4 BD2 mut3 UNOs. h) Representative bright-field microscopy images of 41-day UNOs, Scale bar=1mm (rest of the images in source file). i) Dot plots showing the average expression level (Z scores) and percentage of cells expressed in Glutamatergic, Diencephalic-1(pink in UMAP), and Diencephalic-2(blue in UMAP), and G2M clusters for bivalent genes that showed significant differential expression in the scRNA-seq data in BRD4-BD2 mut1 and BRD4-BD2 mut2 UNOs.

Techniques: Immunofluorescence, Staining, RNA Sequencing, Microscopy, Expressing, Quantitative Proteomics

Journal: bioRxiv

Article Title: BRD4 represses developmental and neuronal genes through interactions with polycomb complexes

doi: 10.64898/2026.01.31.702994

Figure Lengend Snippet: a) UMAP plots show the distribution of single-cell ATAC sequencing (scATAC-seq) data clustered by genotypes WT and BRD4 BD2 mut2 and annotated by cell lineage for WT and BRD4 BD2 mut2. b ) Z-scores (high scores in red and low scores are in blue) showing top transcription factor motifs enriched at Diencephalic, Glutamatergic, G2M and GABAnergic lineages across scATACseq peaks, which are gained in BRD4 BD2 mut 2 UNO compared to WT control. The complete list of enriched TFs is in the source data table.

Techniques: Sequencing, Control

Analysis of correlations with each other among CENP-F and related genes. (A) The PPI network for CENP-F, CDK1, CDK2, CDK7 and BRD4 in human species. (B) The PPI network for CENP-F, CDK1, CDK2, CDK7 and BRD4 in mouse species. (C) The correlation between CENP-F and CDK1 in HCC. (D) The correlation between BRD4 and CDK1 in HCC. (E) The correlation between CENP-F and CDK2 in HCC. (F) The correlation between BRD4 and CDK2 in HCC. (G) The correlation between CENP-F and BRD4 in HCC

Journal: Discover Oncology

Article Title: CENP-F promotes HCC cell proliferation mediated by super enhancer reader BRD4

doi: 10.1007/s12672-025-03785-5

Figure Lengend Snippet: Analysis of correlations with each other among CENP-F and related genes. (A) The PPI network for CENP-F, CDK1, CDK2, CDK7 and BRD4 in human species. (B) The PPI network for CENP-F, CDK1, CDK2, CDK7 and BRD4 in mouse species. (C) The correlation between CENP-F and CDK1 in HCC. (D) The correlation between BRD4 and CDK1 in HCC. (E) The correlation between CENP-F and CDK2 in HCC. (F) The correlation between BRD4 and CDK2 in HCC. (G) The correlation between CENP-F and BRD4 in HCC

Article Snippet: The BRD4 inhibitor JQ1 was purchased from Med Chem Express (HY-13030, Shanghai, China).

Techniques:

The mRNA expression of CENP-F and related genes in HCC. (A) The mRNA expression levels of CENP-F in HCC. (B) The mRNA expression levels of CDK1 in HCC. (C) The mRNA expression levels of CDK2 in HCC. (D) The mRNA expression levels of BRD4 in HCC. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of more than three independent experiments

Journal: Discover Oncology

Article Title: CENP-F promotes HCC cell proliferation mediated by super enhancer reader BRD4

doi: 10.1007/s12672-025-03785-5

Figure Lengend Snippet: The mRNA expression of CENP-F and related genes in HCC. (A) The mRNA expression levels of CENP-F in HCC. (B) The mRNA expression levels of CDK1 in HCC. (C) The mRNA expression levels of CDK2 in HCC. (D) The mRNA expression levels of BRD4 in HCC. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of more than three independent experiments

Article Snippet: The BRD4 inhibitor JQ1 was purchased from Med Chem Express (HY-13030, Shanghai, China).

Techniques: Expressing, Control

The expression of mRNA and protein levels of related genes after knockdown CENP-F and overexpression of CENP-F in vitro. (A) RT-qPCR analysis of CENP-F expression in HepG2 cell line among the normal contral (NC) group, shCENP-F group, empty vector group and overexpression group. (B) RT-qPCR analysis of CDK1 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (C) RT-qPCR analysis of CDK2 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (D) RT-qPCR analysis of BRD4 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (E) RT-qPCR analysis of c-Myc expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (F) RT-qPCR analysis of CENP-F expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (G) RT-qPCR analysis of CDK1 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (H) RT-qPCR analysis of CDK2 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (I) RT-qPCR analysis of BRD4 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (J) RT-qPCR analysis of c-Myc expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. K. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in HepG2 cell line. L. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in Hep3B cell line. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of three independent experiments

Journal: Discover Oncology

Article Title: CENP-F promotes HCC cell proliferation mediated by super enhancer reader BRD4

doi: 10.1007/s12672-025-03785-5

Figure Lengend Snippet: The expression of mRNA and protein levels of related genes after knockdown CENP-F and overexpression of CENP-F in vitro. (A) RT-qPCR analysis of CENP-F expression in HepG2 cell line among the normal contral (NC) group, shCENP-F group, empty vector group and overexpression group. (B) RT-qPCR analysis of CDK1 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (C) RT-qPCR analysis of CDK2 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (D) RT-qPCR analysis of BRD4 expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (E) RT-qPCR analysis of c-Myc expression in HepG2 cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (F) RT-qPCR analysis of CENP-F expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (G) RT-qPCR analysis of CDK1 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (H) RT-qPCR analysis of CDK2 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (I) RT-qPCR analysis of BRD4 expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. (J) RT-qPCR analysis of c-Myc expression in Hep3B cell line among the NC group, shCENP-F group, empty vector group and overexpression group. K. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in HepG2 cell line. L. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in Hep3B cell line. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of three independent experiments

Article Snippet: The BRD4 inhibitor JQ1 was purchased from Med Chem Express (HY-13030, Shanghai, China).

Techniques: Expressing, Knockdown, Over Expression, In Vitro, Quantitative RT-PCR, Plasmid Preparation, Western Blot, Control

Knockdown CENP-F and BRD4 inhibited HCC cell proliferation in vivo. (A) Subcutaneous tumor model of HCC among NC group, shBRD4 and shCENP-F group. (B) Subcutaneous tumor size of HCC among NC group, shBRD4 and shCENP-F group. (C) Tumor growth volume comparison among NC group, shBRD4 and shCENP-F group. (D) Live imaging of tumor tissue after CENP-F inhibition. (E) Live imaging of tumor tissue after BRD4 inhibition. (F) RT-qPCR analysis of CENP-F expression in tumor tissue between the NC group and the shCENP-F group. (G) RT-qPCR analysis of CDK1 expression in tumor tissue between the NC group and the shCENP-F group. (H) RT-qPCR analysis of CDK2 expression in tumor tissue between the NC group and the shCENP-F group. (I) RT-qPCR analysis of BRD4 expression in tumor tissue between the NC group and the shCENP-F group. (J) RT-qPCR analysis of c-Myc expression in tumor tissue between the NC group and the shCENP-F group. K. RT-qPCR analysis of BRD4 expression in tumor tissue between the NC group and the shBRD4 group. L. RT-qPCR analysis of c-Myc expression in tumor tissue between the NC group and the shBRD4 group. M. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in tumor tissue between the NC group and the shCENP-F group. N. Western blotting analysis of BRD4 and c-Myc expression in tumor tissue between the NC group and the shBRD4 group. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of three or six independent experiments

Journal: Discover Oncology

Article Title: CENP-F promotes HCC cell proliferation mediated by super enhancer reader BRD4

doi: 10.1007/s12672-025-03785-5

Figure Lengend Snippet: Knockdown CENP-F and BRD4 inhibited HCC cell proliferation in vivo. (A) Subcutaneous tumor model of HCC among NC group, shBRD4 and shCENP-F group. (B) Subcutaneous tumor size of HCC among NC group, shBRD4 and shCENP-F group. (C) Tumor growth volume comparison among NC group, shBRD4 and shCENP-F group. (D) Live imaging of tumor tissue after CENP-F inhibition. (E) Live imaging of tumor tissue after BRD4 inhibition. (F) RT-qPCR analysis of CENP-F expression in tumor tissue between the NC group and the shCENP-F group. (G) RT-qPCR analysis of CDK1 expression in tumor tissue between the NC group and the shCENP-F group. (H) RT-qPCR analysis of CDK2 expression in tumor tissue between the NC group and the shCENP-F group. (I) RT-qPCR analysis of BRD4 expression in tumor tissue between the NC group and the shCENP-F group. (J) RT-qPCR analysis of c-Myc expression in tumor tissue between the NC group and the shCENP-F group. K. RT-qPCR analysis of BRD4 expression in tumor tissue between the NC group and the shBRD4 group. L. RT-qPCR analysis of c-Myc expression in tumor tissue between the NC group and the shBRD4 group. M. Western blotting analysis of CENP-F, CDK1, CDK2, BRD4 and c-Myc expression in tumor tissue between the NC group and the shCENP-F group. N. Western blotting analysis of BRD4 and c-Myc expression in tumor tissue between the NC group and the shBRD4 group. *<0.05, **<0.01, *** <0.001, ****<0.0001 vs. match control. Statistical significance was tested by Student’s t -test. Data represent mean ± SEM of three or six independent experiments

Article Snippet: The BRD4 inhibitor JQ1 was purchased from Med Chem Express (HY-13030, Shanghai, China).

Techniques: Knockdown, In Vivo, Comparison, Imaging, Inhibition, Quantitative RT-PCR, Expressing, Western Blot, Control

(A) Schematic showing the domain architecture of the Brd4 protein, and how its bromodomains are being used in several combinations to make acyl-eCRs and to determine how the valency of reader domains affects drug perturbations. (B) Immunofluorescence images of mESCs showing the nuclear localization of different valencies of the second bromodomain from BRD4 in the Parbit system (green) and their colocalization with Hoechst (magenta) after drug treatments. All scale bars are 5 µM. Drug treatments were performed at 1 μM concentrations for 24 hours. Bottom panel: Representative pseudocolored images (eGFP signal) depicting the differences in fluorescence intensities in different cell lines. A gradient pseudocolor bar (signal intensity) is shown at the left. (C) Normalized FACS data showing the effects of ARV-825 PROTAC treatment on cells expressing several combinations of bromodomains from BRD4. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells.

Journal: bioRxiv

Article Title: A modular toolbox for in cellulo screening of small molecule inhibitors targeting chromatin reader domains

doi: 10.1101/2025.09.06.674632

Figure Lengend Snippet: (A) Schematic showing the domain architecture of the Brd4 protein, and how its bromodomains are being used in several combinations to make acyl-eCRs and to determine how the valency of reader domains affects drug perturbations. (B) Immunofluorescence images of mESCs showing the nuclear localization of different valencies of the second bromodomain from BRD4 in the Parbit system (green) and their colocalization with Hoechst (magenta) after drug treatments. All scale bars are 5 µM. Drug treatments were performed at 1 μM concentrations for 24 hours. Bottom panel: Representative pseudocolored images (eGFP signal) depicting the differences in fluorescence intensities in different cell lines. A gradient pseudocolor bar (signal intensity) is shown at the left. (C) Normalized FACS data showing the effects of ARV-825 PROTAC treatment on cells expressing several combinations of bromodomains from BRD4. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells.

Article Snippet: The CBP/p300 bromodomain inhibitor: GNE-049 (MedChemExpress, HY-108435), CBP/p300 PROTAC: dCBP-1 (MedChemExpress, HY-134582), BRD4 bromodomain inhibitor: (+)-JQ-1 (MedChemExpress, HY-13030), BRD4 PROTAC: ARV-825 (MedChemExpress, HY-16954), BRD9 bromodomain inhibitor: iBRD9 (MedChemExpress, HY-18975), and broad-spectrum bromodomain inhibitor: Bromosporine (MedChemExpress, HY-15815) were dissolved in DMSO and then diluted to 1μM in mESC media for 24-hour treatments, unless stated otherwise.

Techniques: Immunofluorescence, Fluorescence, Expressing

(A) Top: Schematic showing how the competitive binding of small molecule inhibitors versus PROTACs for the binding pocket of Acyl-eCRs can be used to measure the affinity of a small molecule for a bromodomain in cellulo . Inhibitors with higher affinity for a bromodomain, better prevent PROTAC-induced degradation. Bottom : Treatment scheme for competitive binding experiments. Cells were treated with 1 μM inhibitors for 1 hour. Then, varying concentrations of the PROTAC were added in addition to the previously added inhibitor. After 3 hours of treatment, the cell fluorescence was measured via flow cytometry. (B) Competitive binding between ARV-825 and several small molecule inhibitors showing how the inhibitors bind to BRD4(2)_BRD.1x. The cells were treated with the indicated inhibitor at a 1 μM concentration for 1 hour. Then, the stated concentration of ARV-825 PROTAC was added for 3 hours, in addition to the previous concentration of the same inhibitor. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells. (C) Competitive binding between dCBP-1 and several small molecule inhibitors showing how the inhibitors bind CBP bromodomains in Acyl-eCR constructs versus the endogenous CBP protein. The cells were treated with the indicated inhibitor at a 1 μM concentration for 1 hour. Then, the stated concentration of dCBP-1 PROTAC was added for 3 hours, in addition to the previous concentration of the same inhibitor. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells.

Journal: bioRxiv

Article Title: A modular toolbox for in cellulo screening of small molecule inhibitors targeting chromatin reader domains

doi: 10.1101/2025.09.06.674632

Figure Lengend Snippet: (A) Top: Schematic showing how the competitive binding of small molecule inhibitors versus PROTACs for the binding pocket of Acyl-eCRs can be used to measure the affinity of a small molecule for a bromodomain in cellulo . Inhibitors with higher affinity for a bromodomain, better prevent PROTAC-induced degradation. Bottom : Treatment scheme for competitive binding experiments. Cells were treated with 1 μM inhibitors for 1 hour. Then, varying concentrations of the PROTAC were added in addition to the previously added inhibitor. After 3 hours of treatment, the cell fluorescence was measured via flow cytometry. (B) Competitive binding between ARV-825 and several small molecule inhibitors showing how the inhibitors bind to BRD4(2)_BRD.1x. The cells were treated with the indicated inhibitor at a 1 μM concentration for 1 hour. Then, the stated concentration of ARV-825 PROTAC was added for 3 hours, in addition to the previous concentration of the same inhibitor. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells. (C) Competitive binding between dCBP-1 and several small molecule inhibitors showing how the inhibitors bind CBP bromodomains in Acyl-eCR constructs versus the endogenous CBP protein. The cells were treated with the indicated inhibitor at a 1 μM concentration for 1 hour. Then, the stated concentration of dCBP-1 PROTAC was added for 3 hours, in addition to the previous concentration of the same inhibitor. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells.

Techniques: Binding Assay, Fluorescence, Flow Cytometry, Concentration Assay, Construct

TGF-β-induced BRD4 expression along with SMC markers in 10T1/2 cells. ( A , B ), TGF-β-induced BRD4 and SMC marker expression dose dependently in 10T1/2 cells. Serum-starved 10T1/2 cells were treated with vehicle or different concentrations of TGF-β as indicated for 48 h. BRD4 and VSMC markers (α-SMA and SM22α) were detected by Western blotting (WB, ( A )) and quantified in ( B ). Tubulin served as the loading control. *, p < 0.05 compared to vehicle group (0 ng/mL, ( B )), n = 3 independent experiments. ( C , D ), 10T1/2 cells were starved for 24 h, followed by vehicle or TGF-β (5 ng/mL) induction for various times as indicated. WB ( C ) and qPCR ( D ) were performed to detect BRD4 and SMC marker protein and mRNA expression, respectively. *, p < 0.05 compared to vehicle group (0 h, ( D )), n = 3 replicates. Cyclophilin was the internal control for qPCR.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: TGF-β-induced BRD4 expression along with SMC markers in 10T1/2 cells. ( A , B ), TGF-β-induced BRD4 and SMC marker expression dose dependently in 10T1/2 cells. Serum-starved 10T1/2 cells were treated with vehicle or different concentrations of TGF-β as indicated for 48 h. BRD4 and VSMC markers (α-SMA and SM22α) were detected by Western blotting (WB, ( A )) and quantified in ( B ). Tubulin served as the loading control. *, p < 0.05 compared to vehicle group (0 ng/mL, ( B )), n = 3 independent experiments. ( C , D ), 10T1/2 cells were starved for 24 h, followed by vehicle or TGF-β (5 ng/mL) induction for various times as indicated. WB ( C ) and qPCR ( D ) were performed to detect BRD4 and SMC marker protein and mRNA expression, respectively. *, p < 0.05 compared to vehicle group (0 h, ( D )), n = 3 replicates. Cyclophilin was the internal control for qPCR.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Marker, Western Blot, Control

BRD4 knockdown attenuated TGF-β-induced differentiation of 10T1/2 cells to SMCs. ( A ), 10T1/2 cells were transfected with scramble or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 48 h. The expression of BRD4 and SMC markers was detected by WB ( A ). ( B ) is the quantification of A based on 3 independent experiments. *, p < 0.05 compared to vehicle-treated scramble; #, p < 0.05 compared to TGF-β-treated scramble.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 knockdown attenuated TGF-β-induced differentiation of 10T1/2 cells to SMCs. ( A ), 10T1/2 cells were transfected with scramble or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 48 h. The expression of BRD4 and SMC markers was detected by WB ( A ). ( B ) is the quantification of A based on 3 independent experiments. *, p < 0.05 compared to vehicle-treated scramble; #, p < 0.05 compared to TGF-β-treated scramble.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Knockdown, Transfection, Expressing

BRD4 inhibitor JQ1 inhibited TGF-β-induced differentiation of 10T1/2 cells into SMCs. ( A , B ), serum-starved 10T1/2 cells were pretreated with different doses of JQ1, followed by TGF-β treatment for an additional 48 h, and cells were then harvested for WB analysis of SMC markers. Tubulin was used as the loading control. B is the quantification of A based on 3 independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. ( C ), Serum-starved 10T1/2 cells were pretreated with different doses of JQ1, followed by TGF-β treatment for an additional 16 h before cells were harvested for qPCR analysis of SMC markers. Cyclophilin was used as an internal control. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. n = 3 replicates.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 inhibitor JQ1 inhibited TGF-β-induced differentiation of 10T1/2 cells into SMCs. ( A , B ), serum-starved 10T1/2 cells were pretreated with different doses of JQ1, followed by TGF-β treatment for an additional 48 h, and cells were then harvested for WB analysis of SMC markers. Tubulin was used as the loading control. B is the quantification of A based on 3 independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. ( C ), Serum-starved 10T1/2 cells were pretreated with different doses of JQ1, followed by TGF-β treatment for an additional 16 h before cells were harvested for qPCR analysis of SMC markers. Cyclophilin was used as an internal control. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. n = 3 replicates.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Control

BRD4 degraders inhibited TGF-β-induced differentiation of 10T1/2 cells into SMCs. ( A , C ), Serum-starved 10T1/2 cells were pretreated with different doses of ARV-825 (ARV, ( A )) or dBET1 ( C ), followed by TGF-β treatment (5 ng/mL) for an additional 48 h, and the cells were then harvested for WB analysis of SMC markers. Tubulin was used as the loading control. ( B , D ) are the quantification of ( A , C ), respectively, based on 3 independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 degraders inhibited TGF-β-induced differentiation of 10T1/2 cells into SMCs. ( A , C ), Serum-starved 10T1/2 cells were pretreated with different doses of ARV-825 (ARV, ( A )) or dBET1 ( C ), followed by TGF-β treatment (5 ng/mL) for an additional 48 h, and the cells were then harvested for WB analysis of SMC markers. Tubulin was used as the loading control. ( B , D ) are the quantification of ( A , C ), respectively, based on 3 independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Control

BRD4 knockdown inhibited TGF-β-induced TAZ expression in 10T1/2 cells. ( A , B ), Serum-starved 10T1/2 cells were treated with TGF-β treatment (5 ng/mL) for different times as indicated before the cells were harvested for WB ( A ) and qPCR ( B ) analysis of TAZ. Tubulin was used as the loading control. Cyclophilin was used as the internal control for qPCR analysis in B. *, p < 0.05 compared to control (0 h), n = 3 replicates. ( C ), 10T1/2 cells were transfected with scramble control or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 8 h. The cells were then harvested for analysis of given protein by WB. ( D ) is the quantification of C based on 3 independent experiments. *, p < 0.05 compared to the vehicle-treated scramble; #, p < 0.05 compared to TGF-β-treated scramble.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 knockdown inhibited TGF-β-induced TAZ expression in 10T1/2 cells. ( A , B ), Serum-starved 10T1/2 cells were treated with TGF-β treatment (5 ng/mL) for different times as indicated before the cells were harvested for WB ( A ) and qPCR ( B ) analysis of TAZ. Tubulin was used as the loading control. Cyclophilin was used as the internal control for qPCR analysis in B. *, p < 0.05 compared to control (0 h), n = 3 replicates. ( C ), 10T1/2 cells were transfected with scramble control or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 8 h. The cells were then harvested for analysis of given protein by WB. ( D ) is the quantification of C based on 3 independent experiments. *, p < 0.05 compared to the vehicle-treated scramble; #, p < 0.05 compared to TGF-β-treated scramble.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Knockdown, Expressing, Control, Transfection

BRD4 inhibitors suppressed TGF-β-induced TAZ expression in 10T1/2 cells. ( A ), serum-starved 10T1/2 cells were pretreated with JQ1 (2 µM), dBET1 (100 nM), or ARV-825 (ARV, 100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 8 h. The cells were then harvested for WB analysis of TAZ. Tubulin was used as the loading control. ( B ) is the quantification of A based on 3 independent experiments. *, p < 0.05 compared with vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. ( C – E ), serum-starved 10T1/2 cells were pretreated with different doses of JQ1 ( C ), ARV-825 (ARV, ( D )) or dBET1 ( E ), followed by TGF-β treatment (5 ng/mL) for an additional 8 h to detect TAZ through WB.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 inhibitors suppressed TGF-β-induced TAZ expression in 10T1/2 cells. ( A ), serum-starved 10T1/2 cells were pretreated with JQ1 (2 µM), dBET1 (100 nM), or ARV-825 (ARV, 100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 8 h. The cells were then harvested for WB analysis of TAZ. Tubulin was used as the loading control. ( B ) is the quantification of A based on 3 independent experiments. *, p < 0.05 compared with vehicle control; #, p < 0.05 compared to TGF-β treatment alone group. ( C – E ), serum-starved 10T1/2 cells were pretreated with different doses of JQ1 ( C ), ARV-825 (ARV, ( D )) or dBET1 ( E ), followed by TGF-β treatment (5 ng/mL) for an additional 8 h to detect TAZ through WB.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Control

Myocardin is involved in BRD4 mediated10T1/2 differentiation into SMCs. ( A ) Serum-starved 10T1/2 cells were treated with TGF-β (5 ng/mL) for various times to detect Myocardin expression through WB. ( B ) is the quantification of A based on three independent experiments. *, p < 0.05 compared with control (0 h). ( C ) 10T1/2 cells were transfected with scramble control or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 24 h. Myocardin expression was detected through WB. ( D ) is the quantification of C based on three independent experiments. *, p < 0.05 compared with the vehicle-treated scramble; #, p < 0.05 compared with TGF-β-treated scramble. ( E ) serum-starved 10T1/2 cells were pretreated with JQ1 (2 µM), dBET1 (100 nM), or ARV-825 (100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 24 h. The cells were then harvested for WB analysis of myocardin. ( F ) is the quantification of E based on three independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group.

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: Myocardin is involved in BRD4 mediated10T1/2 differentiation into SMCs. ( A ) Serum-starved 10T1/2 cells were treated with TGF-β (5 ng/mL) for various times to detect Myocardin expression through WB. ( B ) is the quantification of A based on three independent experiments. *, p < 0.05 compared with control (0 h). ( C ) 10T1/2 cells were transfected with scramble control or siBRD4, followed by treatment with vehicle or TGF-β (5 ng/mL) for an additional 24 h. Myocardin expression was detected through WB. ( D ) is the quantification of C based on three independent experiments. *, p < 0.05 compared with the vehicle-treated scramble; #, p < 0.05 compared with TGF-β-treated scramble. ( E ) serum-starved 10T1/2 cells were pretreated with JQ1 (2 µM), dBET1 (100 nM), or ARV-825 (100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 24 h. The cells were then harvested for WB analysis of myocardin. ( F ) is the quantification of E based on three independent experiments. *, p < 0.05 compared to vehicle control; #, p < 0.05 compared to TGF-β treatment alone group.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Control, Transfection

$BRD4 inhibition reduced the nuclear levels of Smad3 induced by TGF-β in 10T1/2 cells. ( A ) Serum-starved 10T1/2 cells were pretreated with ARV-825 (100 nM) for 30 min, followed by TGF-β treatment (5 ng/mL) for an additional 2 h, and cells were then subject to immunofluorescence staining of Smad3. Images were taken at 200×. ( B ) Serum-starved 10T1/2 cells were pretreated with JQ1 (1 µM) or ARV-825 (100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 2 h, and cells were then subject to fractionation WB analysis of Smad3. Tubulin and Lamin B1 were used as loading controls for the cytoplasm and nucleus, respectively.$

Journal: International Journal of Molecular Sciences

Article Title: BRD4 Mediates Transforming Growth Factor-β-Induced Smooth Muscle Cell Differentiation from Mesenchymal Progenitor Cells

doi: 10.3390/ijms26168074

Figure Lengend Snippet: BRD4 inhibition reduced the nuclear levels of Smad3 induced by TGF-β in 10T1/2 cells. ( A ) Serum-starved 10T1/2 cells were pretreated with ARV-825 (100 nM) for 30 min, followed by TGF-β treatment (5 ng/mL) for an additional 2 h, and cells were then subject to immunofluorescence staining of Smad3. Images were taken at 200×. ( B ) Serum-starved 10T1/2 cells were pretreated with JQ1 (1 µM) or ARV-825 (100 nM), followed by TGF-β treatment (5 ng/mL) for an additional 2 h, and cells were then subject to fractionation WB analysis of Smad3. Tubulin and Lamin B1 were used as loading controls for the cytoplasm and nucleus, respectively.

Article Snippet: The small molecular inhibitor of BRD4 JQ1 (HY-13030) and degraders ARV-825 (HY-16954) and dBET1 (HY-101838) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Immunofluorescence, Staining, Fractionation

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1) Product Images from "BRD4 represses developmental and neuronal genes through interactions with polycomb complexes"

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