anti acetyl histone h3  (Cell Signaling Technology Inc)

 
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    Name:
    Histone H3 D2B12 XP Rabbit mAb ChIP Formulated
    Description:
    Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes The nucleosome made up of DNA wound around eight core histone proteins two each of H2A H2B H3 and H4 is the primary building block of chromatin 1 The amino terminal tails of core histones undergo various post translational modifications including acetylation phosphorylation methylation and ubiquitination 2 5 These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and therefore gene expression 6 In most species histone H2B is primarily acetylated at Lys5 12 15 and 20 4 7 Histone H3 is primarily acetylated at Lys9 14 18 23 27 and 56 Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms 2 3 Phosphorylation at Ser10 Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis 8 10 Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin Immunostaining with phospho specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase 11
    Catalog Number:
    4620
    Price:
    None
    Applications:
    Chromatin Immunoprecipitation
    Category:
    Primary Antibodies
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein.
    Reactivity:
    Human Mouse
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc anti acetyl histone h3
    Priming, but not cotreatment, of primary human macrophages with HDAC inhibitors impairs phagocytosis of E. coli . (A) HMDM were treated with 1 μM TSA or 10 μM SAHA for 6 h, after which cell lysates were prepared and <t>histone</t> H3 hyperacetylation
    Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes The nucleosome made up of DNA wound around eight core histone proteins two each of H2A H2B H3 and H4 is the primary building block of chromatin 1 The amino terminal tails of core histones undergo various post translational modifications including acetylation phosphorylation methylation and ubiquitination 2 5 These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and therefore gene expression 6 In most species histone H2B is primarily acetylated at Lys5 12 15 and 20 4 7 Histone H3 is primarily acetylated at Lys9 14 18 23 27 and 56 Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms 2 3 Phosphorylation at Ser10 Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis 8 10 Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin Immunostaining with phospho specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase 11
    https://www.bioz.com/result/anti acetyl histone h3/product/Cell Signaling Technology Inc
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti acetyl histone h3 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Histone Deacetylase Inhibitors Promote Mitochondrial Reactive Oxygen Species Production and Bacterial Clearance by Human Macrophages"

    Article Title: Histone Deacetylase Inhibitors Promote Mitochondrial Reactive Oxygen Species Production and Bacterial Clearance by Human Macrophages

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01876-15

    Priming, but not cotreatment, of primary human macrophages with HDAC inhibitors impairs phagocytosis of E. coli . (A) HMDM were treated with 1 μM TSA or 10 μM SAHA for 6 h, after which cell lysates were prepared and histone H3 hyperacetylation
    Figure Legend Snippet: Priming, but not cotreatment, of primary human macrophages with HDAC inhibitors impairs phagocytosis of E. coli . (A) HMDM were treated with 1 μM TSA or 10 μM SAHA for 6 h, after which cell lysates were prepared and histone H3 hyperacetylation

    Techniques Used:

    2) Product Images from "Toxoplasma gondii Prevents Chromatin Remodeling Initiated by TLR-Triggered Macrophage Activation 1"

    Article Title: Toxoplasma gondii Prevents Chromatin Remodeling Initiated by TLR-Triggered Macrophage Activation 1

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    T. gondii globally blocks LPS-induced phosphorylation of histone H3 at Ser 10 but has no effect on Lys 9/14 histone H3 acetylation
    Figure Legend Snippet: T. gondii globally blocks LPS-induced phosphorylation of histone H3 at Ser 10 but has no effect on Lys 9/14 histone H3 acetylation

    Techniques Used:

    T. gondii blocks LPS-induced histone H3 modification at the TNF promoter
    Figure Legend Snippet: T. gondii blocks LPS-induced histone H3 modification at the TNF promoter

    Techniques Used: Modification

    3) Product Images from "Selective Histone Deacetylase 6 Inhibitor 23BB Alleviated Rhabdomyolysis-Induced Acute Kidney Injury by Regulating Endoplasmic Reticulum Stress and Apoptosis"

    Article Title: Selective Histone Deacetylase 6 Inhibitor 23BB Alleviated Rhabdomyolysis-Induced Acute Kidney Injury by Regulating Endoplasmic Reticulum Stress and Apoptosis

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2018.00274

    23BB inhibits the expression of HDAC6 and enhances the acetylation of histone H3. (A) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies against HDAC6 and acetylated histone H3. (B) Expressions of HDAC6 and acetylated histone H3 were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 6 for each group). ∗ P
    Figure Legend Snippet: 23BB inhibits the expression of HDAC6 and enhances the acetylation of histone H3. (A) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies against HDAC6 and acetylated histone H3. (B) Expressions of HDAC6 and acetylated histone H3 were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 6 for each group). ∗ P

    Techniques Used: Expressing

    4) Product Images from "2-Methylquinazoline derivative 23BB as a highly selective histone deacetylase 6 inhibitor alleviated cisplatin-induced acute kidney injury"

    Article Title: 2-Methylquinazoline derivative 23BB as a highly selective histone deacetylase 6 inhibitor alleviated cisplatin-induced acute kidney injury

    Journal: Bioscience Reports

    doi: 10.1042/BSR20191538

    Inhibition of HDAC6 activity suppressed apoptosis and ER stress in cisplatin-induced renal proximal tubule HK-2 cells ( A ) HK-2 cells were treated with 23BB, subjected to Annexin-V/PI staining, and then analyzed the apoptosis by flow cytometry. ( B ) The expression of HDAC6, acetylated histone H3 and acetylated α-tubulin in HK-2 cells as measured by immunoblot analysis. ( C ) The expression of CHOP, BCL-2, BAX and cleaved caspase 3 in HK-2 cells as measured by immunoblot analysis. ( D ) The expression of GRP78, p-PERK, p-eIF2α/eIF2α and ATF4 in HK-2 cells as measured by immunoblot analysis.
    Figure Legend Snippet: Inhibition of HDAC6 activity suppressed apoptosis and ER stress in cisplatin-induced renal proximal tubule HK-2 cells ( A ) HK-2 cells were treated with 23BB, subjected to Annexin-V/PI staining, and then analyzed the apoptosis by flow cytometry. ( B ) The expression of HDAC6, acetylated histone H3 and acetylated α-tubulin in HK-2 cells as measured by immunoblot analysis. ( C ) The expression of CHOP, BCL-2, BAX and cleaved caspase 3 in HK-2 cells as measured by immunoblot analysis. ( D ) The expression of GRP78, p-PERK, p-eIF2α/eIF2α and ATF4 in HK-2 cells as measured by immunoblot analysis.

    Techniques Used: Inhibition, Activity Assay, Staining, Flow Cytometry, Cytometry, Expressing

    Inhibition of HDAC6 enhanced the acetylation of histone H3 and α-tubulin ( A ) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies. ( B ) Expressions of acetylated histone H3 and α-tubulin were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 3). *** P
    Figure Legend Snippet: Inhibition of HDAC6 enhanced the acetylation of histone H3 and α-tubulin ( A ) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies. ( B ) Expressions of acetylated histone H3 and α-tubulin were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 3). *** P

    Techniques Used: Inhibition

    5) Product Images from "miR-185 mediates lung epithelial cell death after oxidative stress"

    Article Title: miR-185 mediates lung epithelial cell death after oxidative stress

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00392.2015

    Hyperoxia regulates histone acetylation in lung epithelial cells.  A  and  B : Western immunoblot analysis of acetyl-histone H3 ( A ) and acetyl-histone H4 ( B ) protein expressions in human lung epithelial cells (Beas2B) after hyperoxia.  C  and  D : relative acetyl-histone
    Figure Legend Snippet: Hyperoxia regulates histone acetylation in lung epithelial cells. A and B : Western immunoblot analysis of acetyl-histone H3 ( A ) and acetyl-histone H4 ( B ) protein expressions in human lung epithelial cells (Beas2B) after hyperoxia. C and D : relative acetyl-histone

    Techniques Used: Western Blot

    The expression of miR-185 is controlled by HDAC4. A : schematic representation of the genomic location of the human TANGO2 gene. miR-185 locates in the intron of the TANGO2 gene. The acetylation of lysine 27 of the H3 histone protein (H3K27Ac) mark is
    Figure Legend Snippet: The expression of miR-185 is controlled by HDAC4. A : schematic representation of the genomic location of the human TANGO2 gene. miR-185 locates in the intron of the TANGO2 gene. The acetylation of lysine 27 of the H3 histone protein (H3K27Ac) mark is

    Techniques Used: Expressing

    6) Product Images from "DNA METHYLATION AND HISTONE ACETYLATION WORK IN CONCERT TO REGULATE MEMORY FORMATION AND SYNAPTIC PLASTICITY"

    Article Title: DNA METHYLATION AND HISTONE ACETYLATION WORK IN CONCERT TO REGULATE MEMORY FORMATION AND SYNAPTIC PLASTICITY

    Journal: Neurobiology of learning and memory

    doi: 10.1016/j.nlm.2007.07.016

    DNMT inhibition blocks memory consolidation and histone acetylation. (a) Intra-CA1 infusion of 5-AZA immediately following contextual fear conditioning blocked consolidation, as evidenced by a lack of freezing at the 24 h test (N = 7, C/S + VEH; N = 9, C/S + 5-AZA). (b) Intra-CA1 infusion of 5-AZA immediately following contextual fear conditioning (C/S + 5-AZA) blocked AcH3 one hr post-training, but had no effect on context only controls (C + 5-AZA; N = 7 per group). (c) Intra-CA1 infusion of 5-AZA had no effect on the ERK phosphorylation induced by contextual fear conditioning. (N = 7 per group.) * P
    Figure Legend Snippet: DNMT inhibition blocks memory consolidation and histone acetylation. (a) Intra-CA1 infusion of 5-AZA immediately following contextual fear conditioning blocked consolidation, as evidenced by a lack of freezing at the 24 h test (N = 7, C/S + VEH; N = 9, C/S + 5-AZA). (b) Intra-CA1 infusion of 5-AZA immediately following contextual fear conditioning (C/S + 5-AZA) blocked AcH3 one hr post-training, but had no effect on context only controls (C + 5-AZA; N = 7 per group). (c) Intra-CA1 infusion of 5-AZA had no effect on the ERK phosphorylation induced by contextual fear conditioning. (N = 7 per group.) * P

    Techniques Used: Inhibition

    7) Product Images from "Histone deacetylase inhibitor treatment induces ‘BRCAness’ and synergistic lethality with PARP inhibitor and cisplatin against human triple negative breast cancer cells"

    Article Title: Histone deacetylase inhibitor treatment induces ‘BRCAness’ and synergistic lethality with PARP inhibitor and cisplatin against human triple negative breast cancer cells

    Journal: Oncotarget

    doi:

    Treatment with panobinostat induces proteasomal degradation of BRCA1, ATR and CHK1 in breast cancer cells A-B. SUM159PT and HCC1937 cells were treated with 50 nM PS and/or 10 nM carfilzomib (CZ), as indicated, for 24 hours. Following this, cell lysates were prepared and immunoblot analyses were performed for the expression levels of BRCA1, ATR, CHK1, RAD52, KU70, γ-H2AX, acetylated α-tubulin, acetylated histone H3 and β-actin in the cell lysates. The numbers underneath the bands represent densitometry relative to the untreated cells.
    Figure Legend Snippet: Treatment with panobinostat induces proteasomal degradation of BRCA1, ATR and CHK1 in breast cancer cells A-B. SUM159PT and HCC1937 cells were treated with 50 nM PS and/or 10 nM carfilzomib (CZ), as indicated, for 24 hours. Following this, cell lysates were prepared and immunoblot analyses were performed for the expression levels of BRCA1, ATR, CHK1, RAD52, KU70, γ-H2AX, acetylated α-tubulin, acetylated histone H3 and β-actin in the cell lysates. The numbers underneath the bands represent densitometry relative to the untreated cells.

    Techniques Used: Expressing

    Co-treatment with VS and ABT-888 significantly inhibits tumor growth and improves the survival of NOD/SCID mice bearing MDA-MB-231 xenografts A. Mean tumor volume of mice treated with vehicle, ABT-888 and/or VS for 3 weeks. Mice treated with ABT-888 and VS displayed significantly smaller tumors than mice treated with ABT-888 alone (p=0.037) or VS alone (p=0.04). B. Kaplan-Meier survival plot of the mice treated with vehicle, ABT-888, VS, or ABT-888+VS. Mice treated with the combination of ABT-888 and VS demonstrated significantly improved survival (p=0.02) by Log rank (Mantel-Cox) test. C. Representative immunoblots of BRCA1, ATR, CHK1, RAD52, γ-H2AX, cleaved Caspase 3, acetyl histone H3, BIM and β-actin in cell lysates from tumors harvested from mice following 1 week of treatment with ABT-888 and/or VS. D. HCC1937, MDA-MB-231 and SUM159PT cells were treated with cisplatin and vorinostat (VS) for 48 hours and the % apoptotic cells was determined by flow cytometry. Median dose effect and isobologram analyses were performed using Calcusyn. Combination index (CI) values less than 1.0 indicate a synergistic interaction of the two agents in the combination. E. Pan-HDAC inhibitor, by inhibiting HDAC6 levels and activity, induces acetylation and inhibits the chaperone activity of hsp90. This disrupts the chaperone association of hsp90 with its client proteins, such as ATR, BRCA1, RAD52 and CHK1, leading to depletion of their expression levels. Cisplatin treatment leads to decreases in DNA repair and abrogation of cell cycle checkpoints. Treatment with PARP inhibitor inhibits DNA repair leading to accumulation of DNA damage. Combined treatment with HDAC inhibitor and PARP inhibitor leads to greater DNA damage and increased cell death through increased ROS and inhibition of homologous recombination due to depletion of BRCA1 and RAD52. Combined treatment with pan HDAC inhibitor and cisplatin causes greater abrogation of cell cycle checkpoints through HDAC inhibitor-mediated depletion of ATR and CHK1. In addition, accumulation of DNA damage from the combined action of HDAC inhibitor and cisplatin leads to increased cell death of breast cancer cells.
    Figure Legend Snippet: Co-treatment with VS and ABT-888 significantly inhibits tumor growth and improves the survival of NOD/SCID mice bearing MDA-MB-231 xenografts A. Mean tumor volume of mice treated with vehicle, ABT-888 and/or VS for 3 weeks. Mice treated with ABT-888 and VS displayed significantly smaller tumors than mice treated with ABT-888 alone (p=0.037) or VS alone (p=0.04). B. Kaplan-Meier survival plot of the mice treated with vehicle, ABT-888, VS, or ABT-888+VS. Mice treated with the combination of ABT-888 and VS demonstrated significantly improved survival (p=0.02) by Log rank (Mantel-Cox) test. C. Representative immunoblots of BRCA1, ATR, CHK1, RAD52, γ-H2AX, cleaved Caspase 3, acetyl histone H3, BIM and β-actin in cell lysates from tumors harvested from mice following 1 week of treatment with ABT-888 and/or VS. D. HCC1937, MDA-MB-231 and SUM159PT cells were treated with cisplatin and vorinostat (VS) for 48 hours and the % apoptotic cells was determined by flow cytometry. Median dose effect and isobologram analyses were performed using Calcusyn. Combination index (CI) values less than 1.0 indicate a synergistic interaction of the two agents in the combination. E. Pan-HDAC inhibitor, by inhibiting HDAC6 levels and activity, induces acetylation and inhibits the chaperone activity of hsp90. This disrupts the chaperone association of hsp90 with its client proteins, such as ATR, BRCA1, RAD52 and CHK1, leading to depletion of their expression levels. Cisplatin treatment leads to decreases in DNA repair and abrogation of cell cycle checkpoints. Treatment with PARP inhibitor inhibits DNA repair leading to accumulation of DNA damage. Combined treatment with HDAC inhibitor and PARP inhibitor leads to greater DNA damage and increased cell death through increased ROS and inhibition of homologous recombination due to depletion of BRCA1 and RAD52. Combined treatment with pan HDAC inhibitor and cisplatin causes greater abrogation of cell cycle checkpoints through HDAC inhibitor-mediated depletion of ATR and CHK1. In addition, accumulation of DNA damage from the combined action of HDAC inhibitor and cisplatin leads to increased cell death of breast cancer cells.

    Techniques Used: Mouse Assay, Multiple Displacement Amplification, Western Blot, Flow Cytometry, Cytometry, Activity Assay, Expressing, Inhibition, Homologous Recombination

    8) Product Images from "The short-term effect of histone deacetylase inhibitors, chidamide and valproic acid, on the NF-κB pathway in multiple myeloma cells"

    Article Title: The short-term effect of histone deacetylase inhibitors, chidamide and valproic acid, on the NF-κB pathway in multiple myeloma cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3963

    Chidamide and VPA enhance histone H3 acetylation and inhibit HDAC activity in mM cell lines at 48 h, but not at 0 and 6 h. (A) Chidamide and VPA promoted an increase in the levels of histone H3 acetylation in human MM cells (RPMI-8226 and U266). (B) Chidamide and VPA inhibited HDAC activity in human mM cells (RPMI-8226 and U266). * P
    Figure Legend Snippet: Chidamide and VPA enhance histone H3 acetylation and inhibit HDAC activity in mM cell lines at 48 h, but not at 0 and 6 h. (A) Chidamide and VPA promoted an increase in the levels of histone H3 acetylation in human MM cells (RPMI-8226 and U266). (B) Chidamide and VPA inhibited HDAC activity in human mM cells (RPMI-8226 and U266). * P

    Techniques Used: Activity Assay

    9) Product Images from "The Autotaxin–Lysophosphatidic Acid Axis Modulates Histone Acetylation and Gene Expression during Oligodendrocyte Differentiation"

    Article Title: The Autotaxin–Lysophosphatidic Acid Axis Modulates Histone Acetylation and Gene Expression during Oligodendrocyte Differentiation

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0345-15.2015

    In rodent OLG cultures, inhibition of the lysoPLD activity of ATX leads to an increase in nuclear histone acetylation at H3K9. A , B , Representative confocal images of differentiating OLGs treated with vehicle (control, top) or HA130 (bottom) and immunolabeled
    Figure Legend Snippet: In rodent OLG cultures, inhibition of the lysoPLD activity of ATX leads to an increase in nuclear histone acetylation at H3K9. A , B , Representative confocal images of differentiating OLGs treated with vehicle (control, top) or HA130 (bottom) and immunolabeled

    Techniques Used: Inhibition, Activity Assay, Immunolabeling

    In rodent OLG cultures, LPA rescue of ATX–lysoPLD activity inhibition requires the activity of class I HDAC members HDAC1 and HDAC2 but not the class II HDAC member HDAC6. A , Experimental design. Timing of histone deacetylation is marked as described
    Figure Legend Snippet: In rodent OLG cultures, LPA rescue of ATX–lysoPLD activity inhibition requires the activity of class I HDAC members HDAC1 and HDAC2 but not the class II HDAC member HDAC6. A , Experimental design. Timing of histone deacetylation is marked as described

    Techniques Used: Activity Assay, Inhibition

    10) Product Images from "Selective Histone Deacetylase 6 Inhibitor 23BB Alleviated Rhabdomyolysis-Induced Acute Kidney Injury by Regulating Endoplasmic Reticulum Stress and Apoptosis"

    Article Title: Selective Histone Deacetylase 6 Inhibitor 23BB Alleviated Rhabdomyolysis-Induced Acute Kidney Injury by Regulating Endoplasmic Reticulum Stress and Apoptosis

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2018.00274

    23BB inhibits the expression of HDAC6 and enhances the acetylation of histone H3. (A) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies against HDAC6 and acetylated histone H3. (B) Expressions of HDAC6 and acetylated histone H3 were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 6 for each group). ∗ P
    Figure Legend Snippet: 23BB inhibits the expression of HDAC6 and enhances the acetylation of histone H3. (A) The kidney tissue lysates were subjected to immunoblot analysis with indicated antibodies against HDAC6 and acetylated histone H3. (B) Expressions of HDAC6 and acetylated histone H3 were quantified by densitometry and normalized with β-actin. Data are represented as the means ± SE ( n = 6 for each group). ∗ P

    Techniques Used: Expressing

    11) Product Images from "Activation of the Growth-Differentiation Factor 11 Gene by the Histone Deacetylase (HDAC) Inhibitor Trichostatin A and Repression by HDAC3"

    Article Title: Activation of the Growth-Differentiation Factor 11 Gene by the Histone Deacetylase (HDAC) Inhibitor Trichostatin A and Repression by HDAC3

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.24.12.5106-5118.2004

    Enhanced H3K9 acetylation on the gdf11 promoter upon TSA treatment or HDAC3 silencing. (A) Core histones prepared from HeLa cells treated with 400 ng of TSA/ml (left panels) or transfected with plasmids expressing siRNAs (right panels) were Western blotted with antibodies against acetylated H3, H3K9, H3K18, H4, H4K8, and H4K12. A Coomassie-stained gel was prepared in parallel to assess the quality of each histone preparation. (B) Schematic diagram of the human gdf11 promoter (top panel). The arrows facing in opposite directions above the promoter denote the primers used in the ChIP assays. Cross-linked chromatin prepared from HeLa cells treated with (+) or without (−) TSA (middle panels) or transfected with siRNA expression plasmids (bottom panel) was precipitated with the indicated antibodies, and PCR analysis was performed using primers specific for the gdf11 promoter region.
    Figure Legend Snippet: Enhanced H3K9 acetylation on the gdf11 promoter upon TSA treatment or HDAC3 silencing. (A) Core histones prepared from HeLa cells treated with 400 ng of TSA/ml (left panels) or transfected with plasmids expressing siRNAs (right panels) were Western blotted with antibodies against acetylated H3, H3K9, H3K18, H4, H4K8, and H4K12. A Coomassie-stained gel was prepared in parallel to assess the quality of each histone preparation. (B) Schematic diagram of the human gdf11 promoter (top panel). The arrows facing in opposite directions above the promoter denote the primers used in the ChIP assays. Cross-linked chromatin prepared from HeLa cells treated with (+) or without (−) TSA (middle panels) or transfected with siRNA expression plasmids (bottom panel) was precipitated with the indicated antibodies, and PCR analysis was performed using primers specific for the gdf11 promoter region.

    Techniques Used: Transfection, Expressing, Western Blot, Staining, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    12) Product Images from "Histone variant Htz1 promotes histone H3 acetylation to enhance nucleotide excision repair in Htz1 nucleosomes"

    Article Title: Histone variant Htz1 promotes histone H3 acetylation to enhance nucleotide excision repair in Htz1 nucleosomes

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkt688

    The occupancy of Gcn5 and induction of H3 acetylation at the MFA2 promoter is partially dependent on Htz1. ( A ) Gcn5 expression in wild-type and the htz1Δ mutant. Gcn5 was detected with anti-Myc antibodies and Cy5 conjugated anti-mouse IgG secondary antibodies in GCN5 -myc strains. The expression levels were normalized against those of Actin. ( B ) ChIP analysis of the occupancy of Gcn5 was performed with anti-Myc antibodies in GCN5 -myc strains. The levels of Gcn5 binding at MFA2 was normalized against those at the HMRa1 locus, and then presented as the fold change relative to the mock irradiated sample in wild type. ( C ) ChIP analysis of Histone H3 acetylation (H3-Ac) was performed using Ac-H3 (K9, K14) antibodies. The H3 acetylation levels were first normalized against the H3 levels, and then presented as the fold change relative to the mock irradiated sample in wild type. −UV: mock irradiated samples; 0: cells received 100 J/m 2 of UV without repair; 0.5 h repair or 1 h repair: cells were irradiated with UV and then were allowed to repair in YPD for the number of hours indicated. Data are the average of at least three independent experiments ± SD.
    Figure Legend Snippet: The occupancy of Gcn5 and induction of H3 acetylation at the MFA2 promoter is partially dependent on Htz1. ( A ) Gcn5 expression in wild-type and the htz1Δ mutant. Gcn5 was detected with anti-Myc antibodies and Cy5 conjugated anti-mouse IgG secondary antibodies in GCN5 -myc strains. The expression levels were normalized against those of Actin. ( B ) ChIP analysis of the occupancy of Gcn5 was performed with anti-Myc antibodies in GCN5 -myc strains. The levels of Gcn5 binding at MFA2 was normalized against those at the HMRa1 locus, and then presented as the fold change relative to the mock irradiated sample in wild type. ( C ) ChIP analysis of Histone H3 acetylation (H3-Ac) was performed using Ac-H3 (K9, K14) antibodies. The H3 acetylation levels were first normalized against the H3 levels, and then presented as the fold change relative to the mock irradiated sample in wild type. −UV: mock irradiated samples; 0: cells received 100 J/m 2 of UV without repair; 0.5 h repair or 1 h repair: cells were irradiated with UV and then were allowed to repair in YPD for the number of hours indicated. Data are the average of at least three independent experiments ± SD.

    Techniques Used: Expressing, Mutagenesis, Chromatin Immunoprecipitation, Binding Assay, Irradiation

    Model for chromatin remodelling at MFA2 during NER. Top panel. In the absence of UV histone H3 tails remain unacetylated and chromatin remains repressive. Middle Panel. After UV irradiation, the increased occupancy of Gcn5 that is mediated via the Rad7/Rad16 GG-NER complex is promoted by the presence of Htz1. This helps achieve enhanced acetylation levels on histone H3, which are critical for the specific chromatin setting required for optimal NER, so ensuring maximal binding of repair proteins, e.g. Rad14, to damaged DNA. This leads to efficient lesion removal. Lower Panel. Without Htz1 in these nucleosomes, the occupancy of Gcn5 and the acetylation levels on histone H3 are reduced. Therefore, the maximal binding of repair proteins to damaged DNA cannot be achieved and more CPDs remain unrepaired.
    Figure Legend Snippet: Model for chromatin remodelling at MFA2 during NER. Top panel. In the absence of UV histone H3 tails remain unacetylated and chromatin remains repressive. Middle Panel. After UV irradiation, the increased occupancy of Gcn5 that is mediated via the Rad7/Rad16 GG-NER complex is promoted by the presence of Htz1. This helps achieve enhanced acetylation levels on histone H3, which are critical for the specific chromatin setting required for optimal NER, so ensuring maximal binding of repair proteins, e.g. Rad14, to damaged DNA. This leads to efficient lesion removal. Lower Panel. Without Htz1 in these nucleosomes, the occupancy of Gcn5 and the acetylation levels on histone H3 are reduced. Therefore, the maximal binding of repair proteins to damaged DNA cannot be achieved and more CPDs remain unrepaired.

    Techniques Used: Irradiation, Binding Assay

    13) Product Images from "Phase II trial of the histone deacetylase inhibitor Belinostat in women with platinum resistant epithelial ovarian cancer and micropapillary (LMP) ovarian tumors"

    Article Title: Phase II trial of the histone deacetylase inhibitor Belinostat in women with platinum resistant epithelial ovarian cancer and micropapillary (LMP) ovarian tumors

    Journal: European journal of cancer (Oxford, England : 1990)

    doi: 10.1016/j.ejca.2010.02.047

    Acetylated histone H3 measured prior to and following Belinostat (cycle 1). For patient 6 the average mean intensity in tumor was pre 19 units and post 37 units, for stroma pre 22 units and post 32 units. For patient 7 in tumor pre 10 units and post 63 units and stroma pre 14 units and post 49 units
    Figure Legend Snippet: Acetylated histone H3 measured prior to and following Belinostat (cycle 1). For patient 6 the average mean intensity in tumor was pre 19 units and post 37 units, for stroma pre 22 units and post 32 units. For patient 7 in tumor pre 10 units and post 63 units and stroma pre 14 units and post 49 units

    Techniques Used:

    Histone acetylation evaluated using Western Blotting for histone H3 and H4 isolated from PBMCs Acetylated histones detected using anti-Acetyl-Histone H3 or H4 (Lys9,Lys8) antibodies (Cell Signaling Technology,Inc) in PBMCs taken prior to and following Belinostat in patients with Micropapillary/LMP tumors.
    Figure Legend Snippet: Histone acetylation evaluated using Western Blotting for histone H3 and H4 isolated from PBMCs Acetylated histones detected using anti-Acetyl-Histone H3 or H4 (Lys9,Lys8) antibodies (Cell Signaling Technology,Inc) in PBMCs taken prior to and following Belinostat in patients with Micropapillary/LMP tumors.

    Techniques Used: Western Blot, Isolation

    14) Product Images from "Cotylenin A inhibits cell proliferation and induces apoptosis and PAX6 mRNA transcripts in retinoblastoma cell lines"

    Article Title: Cotylenin A inhibits cell proliferation and induces apoptosis and PAX6 mRNA transcripts in retinoblastoma cell lines

    Journal: Molecular Vision

    doi:

    The accumulation of acetylated histone H3 and H4 protein in retinoblastoma cell lines. Western blot analysis of acetylated histone H3 and H4 (acetyl-H3 and H4) protein levels in retinoblastoma cells treated with (+) or without (–) cotylenin A (CN-A) for 5 days. β-Actin was used as a loading control.
    Figure Legend Snippet: The accumulation of acetylated histone H3 and H4 protein in retinoblastoma cell lines. Western blot analysis of acetylated histone H3 and H4 (acetyl-H3 and H4) protein levels in retinoblastoma cells treated with (+) or without (–) cotylenin A (CN-A) for 5 days. β-Actin was used as a loading control.

    Techniques Used: Western Blot

    15) Product Images from "Transcriptional corepressor SHP recruits SIRT1 histone deacetylase to inhibit LRH-1 transactivation"

    Article Title: Transcriptional corepressor SHP recruits SIRT1 histone deacetylase to inhibit LRH-1 transactivation

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkq227

    SHP recruits SIRT1 deacetylase to inhibit LRH1-mediated target gene activation. ( A ) The recruitment of SIRT1 by SHP on CYP7A1 (left) and SHP (right) gene promoters is associated with template-associated histone (H3 and H4) deacetylation. HepG2 cells were treated with indicated adenovirus vectors (50 MOI) for 36–72 h. Twelve hours prior to preparation of cell lysates for ChIP assay cells were treated with NAM (20 µM). Chromatin fragments were prepared and immumoprecipitated with the indicated specific antibodies. DNA fragments covering BARE-I and BARE-II element on CYP7A1 (left) and LRH1-binding regions on SHP promoter (right) were PCR-amplified as described in the ‘Materials and Methods’ section. ( B ) HepG2 cells were infected with adenovirus vectors as indicated for 36–72 h and for the last 12 h cells were treated with NAM (20 µM) as indicated. Media was collected for total bile synthesis using Sep-Pak cartridges as described in the ‘Materials and Methods’ section. Data is representative of atleast three independently performed experiments and shown as mean ± SD; * P
    Figure Legend Snippet: SHP recruits SIRT1 deacetylase to inhibit LRH1-mediated target gene activation. ( A ) The recruitment of SIRT1 by SHP on CYP7A1 (left) and SHP (right) gene promoters is associated with template-associated histone (H3 and H4) deacetylation. HepG2 cells were treated with indicated adenovirus vectors (50 MOI) for 36–72 h. Twelve hours prior to preparation of cell lysates for ChIP assay cells were treated with NAM (20 µM). Chromatin fragments were prepared and immumoprecipitated with the indicated specific antibodies. DNA fragments covering BARE-I and BARE-II element on CYP7A1 (left) and LRH1-binding regions on SHP promoter (right) were PCR-amplified as described in the ‘Materials and Methods’ section. ( B ) HepG2 cells were infected with adenovirus vectors as indicated for 36–72 h and for the last 12 h cells were treated with NAM (20 µM) as indicated. Media was collected for total bile synthesis using Sep-Pak cartridges as described in the ‘Materials and Methods’ section. Data is representative of atleast three independently performed experiments and shown as mean ± SD; * P

    Techniques Used: Histone Deacetylase Assay, Activation Assay, Chromatin Immunoprecipitation, Binding Assay, Polymerase Chain Reaction, Amplification, Infection

    16) Product Images from "Induction of Apoptosis in Intestinal Toxicity to a Histone Deacetylase Inhibitor in a Phase I Study with Pelvic Radiotherapy"

    Article Title: Induction of Apoptosis in Intestinal Toxicity to a Histone Deacetylase Inhibitor in a Phase I Study with Pelvic Radiotherapy

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    doi: 10.4143/crt.2016.080

    Immunoblot analysis of histone H3 acetylation (Ac-H3) and poly-(ADP-ribose) polymerase (PARP) cleavage in human colorectal carcinoma cells (HCT-116 and HT29), human BJ fibroblasts, and rat intestinal epithelial cells (IEC-6) following suberoylanilide hydroxamic acid (SAHA) treatment; Amido Black total protein staining as loading controls.
    Figure Legend Snippet: Immunoblot analysis of histone H3 acetylation (Ac-H3) and poly-(ADP-ribose) polymerase (PARP) cleavage in human colorectal carcinoma cells (HCT-116 and HT29), human BJ fibroblasts, and rat intestinal epithelial cells (IEC-6) following suberoylanilide hydroxamic acid (SAHA) treatment; Amido Black total protein staining as loading controls.

    Techniques Used: Staining

    In vivo intestinal effects of suberoylanilide hydroxamic acid (SAHA) in mice treated for five consecutive days and sacrificed on day 5, 3 hours after the oral administration, in terms of histone H3 acetylation (Ac-H3), poly-(ADP-ribose) polymerase (PARP), and heat shock protein 70 (Hsp70) protein expression, verified by immunoblot analysis with α-tubulin expression as a loading control (shown for two mice in each treatment group) (A), body weight change relative to the start of experiment on day 1 (group average values with standard error of the mean; * p
    Figure Legend Snippet: In vivo intestinal effects of suberoylanilide hydroxamic acid (SAHA) in mice treated for five consecutive days and sacrificed on day 5, 3 hours after the oral administration, in terms of histone H3 acetylation (Ac-H3), poly-(ADP-ribose) polymerase (PARP), and heat shock protein 70 (Hsp70) protein expression, verified by immunoblot analysis with α-tubulin expression as a loading control (shown for two mice in each treatment group) (A), body weight change relative to the start of experiment on day 1 (group average values with standard error of the mean; * p

    Techniques Used: In Vivo, Mouse Assay, Expressing

    17) Product Images from "The short-term effect of histone deacetylase inhibitors, chidamide and valproic acid, on the NF-κB pathway in multiple myeloma cells"

    Article Title: The short-term effect of histone deacetylase inhibitors, chidamide and valproic acid, on the NF-κB pathway in multiple myeloma cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3963

    Chidamide and VPA enhance histone H3 acetylation and inhibit HDAC activity in mM cell lines at 48 h, but not at 0 and 6 h. (A) Chidamide and VPA promoted an increase in the levels of histone H3 acetylation in human MM cells (RPMI-8226 and U266). (B) Chidamide and VPA inhibited HDAC activity in human mM cells (RPMI-8226 and U266). * P
    Figure Legend Snippet: Chidamide and VPA enhance histone H3 acetylation and inhibit HDAC activity in mM cell lines at 48 h, but not at 0 and 6 h. (A) Chidamide and VPA promoted an increase in the levels of histone H3 acetylation in human MM cells (RPMI-8226 and U266). (B) Chidamide and VPA inhibited HDAC activity in human mM cells (RPMI-8226 and U266). * P

    Techniques Used: Activity Assay

    18) Product Images from "Toxoplasma gondii Prevents Chromatin Remodeling Initiated by TLR-Triggered Macrophage Activation 1"

    Article Title: Toxoplasma gondii Prevents Chromatin Remodeling Initiated by TLR-Triggered Macrophage Activation 1

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    T. gondii globally blocks LPS-induced phosphorylation of histone H3 at Ser 10 but has no effect on Lys 9/14 histone H3 acetylation
    Figure Legend Snippet: T. gondii globally blocks LPS-induced phosphorylation of histone H3 at Ser 10 but has no effect on Lys 9/14 histone H3 acetylation

    Techniques Used:

    T. gondii blocks LPS-induced histone H3 modification at the TNF promoter
    Figure Legend Snippet: T. gondii blocks LPS-induced histone H3 modification at the TNF promoter

    Techniques Used: Modification

    19) Product Images from "Requirement of Retinoic Acid Receptor ? for Genipin Derivative-Induced Optic Nerve Regeneration in Adult Rat Retina"

    Article Title: Requirement of Retinoic Acid Receptor ? for Genipin Derivative-Induced Optic Nerve Regeneration in Adult Rat Retina

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071252

    S-Nitrosylation of HDAC2 and the acetylation of histone H3 in the rat retina by IPRG001. (A) S-Nitrosylation of HDAC2 1 day after intraocular administration of IPRG001. NO scavenger, c-PTIO or nNOS inhibitor, ETPI was treated 1 h before IPRG001 treatment. Biotinylated proteins were mixed with avidin beads, eluted and analyzed by western blotting with anti-HDAC2 antibody. **P
    Figure Legend Snippet: S-Nitrosylation of HDAC2 and the acetylation of histone H3 in the rat retina by IPRG001. (A) S-Nitrosylation of HDAC2 1 day after intraocular administration of IPRG001. NO scavenger, c-PTIO or nNOS inhibitor, ETPI was treated 1 h before IPRG001 treatment. Biotinylated proteins were mixed with avidin beads, eluted and analyzed by western blotting with anti-HDAC2 antibody. **P

    Techniques Used: Avidin-Biotin Assay, Western Blot

    20) Product Images from "Sensitizing mucoepidermoid carcinomas to chemotherapy by targeted disruption of cancer stem cells"

    Article Title: Sensitizing mucoepidermoid carcinomas to chemotherapy by targeted disruption of cancer stem cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9884

    Levels of ac. Histone H3 in salivary glands and MEC A. Representative expression of ac. histone H3 in acinar cells (A_#1), intercalated duct (A_#2) and secretory ducts (A_#3) of salivary glands. B. MEC has mixed levels of ac. histone H3, with epidermoid and intermediate cells showing both positive and negative staining; in addition, mucous-like tumor cells negative for ac. histone H3 are found next to positive epidermoid tumor cells. C. Quantification of ac. histone H3 using tumor tissue reveals increased acetylation in squamous and intermediate tumor cells (red) compared to mucous-like tumor cells (green). D. Western blot analysis of baseline expression of ac. histone H3 in UM-HMC1, UM-HMC2, UM-HMC3A, UM-HMC3B, and UM-HMC5 tumor cells. GAPDH served as a loading control.
    Figure Legend Snippet: Levels of ac. Histone H3 in salivary glands and MEC A. Representative expression of ac. histone H3 in acinar cells (A_#1), intercalated duct (A_#2) and secretory ducts (A_#3) of salivary glands. B. MEC has mixed levels of ac. histone H3, with epidermoid and intermediate cells showing both positive and negative staining; in addition, mucous-like tumor cells negative for ac. histone H3 are found next to positive epidermoid tumor cells. C. Quantification of ac. histone H3 using tumor tissue reveals increased acetylation in squamous and intermediate tumor cells (red) compared to mucous-like tumor cells (green). D. Western blot analysis of baseline expression of ac. histone H3 in UM-HMC1, UM-HMC2, UM-HMC3A, UM-HMC3B, and UM-HMC5 tumor cells. GAPDH served as a loading control.

    Techniques Used: Expressing, Negative Staining, Western Blot

    21) Product Images from "Orphan Nuclear Receptor Err? Induces C-Reactive Protein Gene Expression through Induction of ER-Bound Bzip Transmembrane Transcription Factor CREBH"

    Article Title: Orphan Nuclear Receptor Err? Induces C-Reactive Protein Gene Expression through Induction of ER-Bound Bzip Transmembrane Transcription Factor CREBH

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086342

    ERRγ regulates activation of CREBH gene promoter. (A), PGC1α-dependent activation of the mouse CREBH promoter by ERRγ. Transient transfection was performed in 293T cells with the indicated plasmid DNAs. (B), Deletion constructs of the CREBH promoter demonstrate the ERRγ binding site in 293T cells. Transient transfection was performed in 293T cells with the indicated plasmid DNAs. (C), ERRE-dependent activation of the CREBH promoter in 293T cells. 293T cells were transfected with the wild-type or ERRE-mutant CREBH promoter along with ERRγ plasmid DNAs. (D), ChIP assay shows the binding of ERRγ and PGC1α to the endogenous CREBH promoter by semiquantitative PCR. AML12 cells were treated with DMSO or Tm (5µg/mL) for 12 hr. After completion of the treatment, chromatin fragments were prepared and immunoprecipitated with ERRγ, PGC1α, or IgG control antibodies. DNA fragments covering –766 to –642 and –195 to –87 elements on the CREBH promoter were PCR amplified. 10% of the soluble chromatin was used as input. (E), ChIP assay for detection of histone acetylation at the ERRγ/PGC1α binding site under the indicated conditions in AML12 cells. Chromatin fragments were prepared and immunoprecipitated with Acetyl-Histone 3 and Acetyl-Histone 4 antibodies. DNA fragments covering –195 to –87 element on the CREBH promoter were qPCR-amplified as described in the ‘Materials and Methods’ section. Data are representative of three independently performed experiments and shown as mean±SD; *P
    Figure Legend Snippet: ERRγ regulates activation of CREBH gene promoter. (A), PGC1α-dependent activation of the mouse CREBH promoter by ERRγ. Transient transfection was performed in 293T cells with the indicated plasmid DNAs. (B), Deletion constructs of the CREBH promoter demonstrate the ERRγ binding site in 293T cells. Transient transfection was performed in 293T cells with the indicated plasmid DNAs. (C), ERRE-dependent activation of the CREBH promoter in 293T cells. 293T cells were transfected with the wild-type or ERRE-mutant CREBH promoter along with ERRγ plasmid DNAs. (D), ChIP assay shows the binding of ERRγ and PGC1α to the endogenous CREBH promoter by semiquantitative PCR. AML12 cells were treated with DMSO or Tm (5µg/mL) for 12 hr. After completion of the treatment, chromatin fragments were prepared and immunoprecipitated with ERRγ, PGC1α, or IgG control antibodies. DNA fragments covering –766 to –642 and –195 to –87 elements on the CREBH promoter were PCR amplified. 10% of the soluble chromatin was used as input. (E), ChIP assay for detection of histone acetylation at the ERRγ/PGC1α binding site under the indicated conditions in AML12 cells. Chromatin fragments were prepared and immunoprecipitated with Acetyl-Histone 3 and Acetyl-Histone 4 antibodies. DNA fragments covering –195 to –87 element on the CREBH promoter were qPCR-amplified as described in the ‘Materials and Methods’ section. Data are representative of three independently performed experiments and shown as mean±SD; *P

    Techniques Used: Activation Assay, Transfection, Plasmid Preparation, Construct, Binding Assay, Mutagenesis, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Immunoprecipitation, Amplification, Real-time Polymerase Chain Reaction

    22) Product Images from "Preclinical anti-myeloma activity of EDO-S101, a new bendamustine-derived molecule with added HDACi activity, through potent DNA damage induction and impairment of DNA repair"

    Article Title: Preclinical anti-myeloma activity of EDO-S101, a new bendamustine-derived molecule with added HDACi activity, through potent DNA damage induction and impairment of DNA repair

    Journal: Journal of Hematology & Oncology

    doi: 10.1186/s13045-017-0495-y

    EDO-S101 induces potent DNA damage and histone acetylation. a Dose and time response of different proteins implicated in the DNA damage response pathway analyzed on the MM1S cell line. b Dose response of different proteins implicated in the DNA damage response pathway on RPMI-8226 and JJN3 cell lines. c Comet assay on the MM1S cell line after treatment with EDO-S101 as compared with untreated cells. Images are representatives of at least 20 captures performed in two independent experiments. Mean tail moment was calculated at different time points using the OpenComet software. d Dose response (48 h) of acetylated proteins after EDO-S101 treatment in MM1S, RPMI-8226, and JJN3 cell lines. e Dose response of proteins implicated in DNA damage repair and acetylation after 48 h of MM1S treatment with EDO-S101 or bendamustine
    Figure Legend Snippet: EDO-S101 induces potent DNA damage and histone acetylation. a Dose and time response of different proteins implicated in the DNA damage response pathway analyzed on the MM1S cell line. b Dose response of different proteins implicated in the DNA damage response pathway on RPMI-8226 and JJN3 cell lines. c Comet assay on the MM1S cell line after treatment with EDO-S101 as compared with untreated cells. Images are representatives of at least 20 captures performed in two independent experiments. Mean tail moment was calculated at different time points using the OpenComet software. d Dose response (48 h) of acetylated proteins after EDO-S101 treatment in MM1S, RPMI-8226, and JJN3 cell lines. e Dose response of proteins implicated in DNA damage repair and acetylation after 48 h of MM1S treatment with EDO-S101 or bendamustine

    Techniques Used: Single Cell Gel Electrophoresis, Software

    23) Product Images from "Pharmacological Inhibition of Class IIA HDACs by LMK-235 in Pancreatic Neuroendocrine Tumor Cells"

    Article Title: Pharmacological Inhibition of Class IIA HDACs by LMK-235 in Pancreatic Neuroendocrine Tumor Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19103128

    (Acetyl-) Histone H3 and histone deacetylase-5 protein expression following LMK-235 treatment. ( A ) Levels of acetyl-histone H3, histone H3, and HDAC5 were evaluated by immunoblotting after LMK-235 incubation for 24 h, as shown by one representative example blot. β-actin was used as a loading control for each sample. The β-actin expression corresponding to the HDAC5 blot is not shown. ( B , C ): Bars represent the semi-quantitative expression as mean values ± SEM ( n = 3) of band intensities of BON-1 ( B ) and QGP-1 ( C ) cells. Abbreviations: UTC = untreated control.
    Figure Legend Snippet: (Acetyl-) Histone H3 and histone deacetylase-5 protein expression following LMK-235 treatment. ( A ) Levels of acetyl-histone H3, histone H3, and HDAC5 were evaluated by immunoblotting after LMK-235 incubation for 24 h, as shown by one representative example blot. β-actin was used as a loading control for each sample. The β-actin expression corresponding to the HDAC5 blot is not shown. ( B , C ): Bars represent the semi-quantitative expression as mean values ± SEM ( n = 3) of band intensities of BON-1 ( B ) and QGP-1 ( C ) cells. Abbreviations: UTC = untreated control.

    Techniques Used: Histone Deacetylase Assay, Expressing, Incubation

    Acetyl-H3 immunofluorescence following LMK-235 treatment. ( A ) Results of one representative acetylated H3 immunofluorescence staining is shown for a LMK-235 dilution series for both cell lines. All samples were visualized using the same exposure time. Acetylated histone H3 is stained in green and the scale bar indicates 100 µm. Magnification 100×. ( B ) Quantitative immunofluorescence intensities are represented as bars for mean ± SEM. * indicate p
    Figure Legend Snippet: Acetyl-H3 immunofluorescence following LMK-235 treatment. ( A ) Results of one representative acetylated H3 immunofluorescence staining is shown for a LMK-235 dilution series for both cell lines. All samples were visualized using the same exposure time. Acetylated histone H3 is stained in green and the scale bar indicates 100 µm. Magnification 100×. ( B ) Quantitative immunofluorescence intensities are represented as bars for mean ± SEM. * indicate p

    Techniques Used: Immunofluorescence, Staining

    24) Product Images from "Activation of the Growth-Differentiation Factor 11 Gene by the Histone Deacetylase (HDAC) Inhibitor Trichostatin A and Repression by HDAC3"

    Article Title: Activation of the Growth-Differentiation Factor 11 Gene by the Histone Deacetylase (HDAC) Inhibitor Trichostatin A and Repression by HDAC3

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.24.12.5106-5118.2004

    Enhanced H3K9 acetylation on the gdf11 promoter upon TSA treatment or HDAC3 silencing. (A) Core histones prepared from HeLa cells treated with 400 ng of TSA/ml (left panels) or transfected with plasmids expressing siRNAs (right panels) were Western blotted with antibodies against acetylated H3, H3K9, H3K18, H4, H4K8, and H4K12. A Coomassie-stained gel was prepared in parallel to assess the quality of each histone preparation. (B) Schematic diagram of the human gdf11 promoter (top panel). The arrows facing in opposite directions above the promoter denote the primers used in the ChIP assays. Cross-linked chromatin prepared from HeLa cells treated with (+) or without (−) TSA (middle panels) or transfected with siRNA expression plasmids (bottom panel) was precipitated with the indicated antibodies, and PCR analysis was performed using primers specific for the gdf11 promoter region.
    Figure Legend Snippet: Enhanced H3K9 acetylation on the gdf11 promoter upon TSA treatment or HDAC3 silencing. (A) Core histones prepared from HeLa cells treated with 400 ng of TSA/ml (left panels) or transfected with plasmids expressing siRNAs (right panels) were Western blotted with antibodies against acetylated H3, H3K9, H3K18, H4, H4K8, and H4K12. A Coomassie-stained gel was prepared in parallel to assess the quality of each histone preparation. (B) Schematic diagram of the human gdf11 promoter (top panel). The arrows facing in opposite directions above the promoter denote the primers used in the ChIP assays. Cross-linked chromatin prepared from HeLa cells treated with (+) or without (−) TSA (middle panels) or transfected with siRNA expression plasmids (bottom panel) was precipitated with the indicated antibodies, and PCR analysis was performed using primers specific for the gdf11 promoter region.

    Techniques Used: Transfection, Expressing, Western Blot, Staining, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    25) Product Images from "Epigenetic Modification Prevents Excessive Wound Healing and Scar Formation After Glaucoma Filtration Surgery"

    Article Title: Epigenetic Modification Prevents Excessive Wound Healing and Scar Formation After Glaucoma Filtration Surgery

    Journal: Investigative Ophthalmology & Visual Science

    doi: 10.1167/iovs.15-18750

    Suberoylanilide hydroxamic acid treatment increases acetylation of histones in human corneal fibroblasts and rabbit conjunctiva tissues. ( A ) Human corneal fibroblast cells treated with SAHA (2.5 μM) for indicated times and ( B ) rabbit conjunctiva injected with SAHA (50 μM) for indicated times were analyzed by Western blot using anti–Ac-histone H3, anti–Ac-histone H4, and anti–β-actin antibodies. Corresponding Western blot quantitative analysis data provided in  C ,  D ,  E  and  F  panels.
    Figure Legend Snippet: Suberoylanilide hydroxamic acid treatment increases acetylation of histones in human corneal fibroblasts and rabbit conjunctiva tissues. ( A ) Human corneal fibroblast cells treated with SAHA (2.5 μM) for indicated times and ( B ) rabbit conjunctiva injected with SAHA (50 μM) for indicated times were analyzed by Western blot using anti–Ac-histone H3, anti–Ac-histone H4, and anti–β-actin antibodies. Corresponding Western blot quantitative analysis data provided in C , D , E and F panels.

    Techniques Used: Injection, Western Blot

    26) Product Images from "Small Molecule Inhibition of cAMP Response Element Binding Protein in Human Acute Myeloid Leukemia Cells"

    Article Title: Small Molecule Inhibition of cAMP Response Element Binding Protein in Human Acute Myeloid Leukemia Cells

    Journal: Leukemia

    doi: 10.1038/leu.2016.139

    Specificity of CREB Inhibition A) The consensus sequence obtained following CREB ChIP-Seq, mapped against the canonical CRE element sequence. B) Relationship between H3K27 acetylation and CREB-driven gene expression. RNA-Seq, CREB ChIP-Seq and H3K27 histone acetylation-Seq were performed on KG-1 cells treated with 5 μM XX-650-23 or 0.1% DMSO. Combined analysis defined the set of genes bound by CREB in AML cells, and alterations in transcription and histone acetylation occurring secondary to XX-650-23 at those loci. The percent of histone acetylation remaining following treatment with XX-650-23 is plotted versus the corresponding change in gene expression for all CREB-bound genes identified by CREB ChIP-Seq. C) Changes in H3K27 acetylation for CREB-bound and -unbound genes were averaged and plotted against base-pair distance to transcriptional start sites (TSS) following XX-650-23 or DMSO treatment. Non-CREB-bound genes show no significant change in H3K27 acetylation following XX-650-23 treatment. D) Heatmap of CREB binding and H3K27 acetylation relative to TSS in DMSO and XX-650-23-treated samples. H3K27 signal intensity, but not CREB binding signal intensity, decreased following XX-650-23. E) Western blot of total and H3K27-specific histone acetylation following XX-650-23 treatment following 6 or 24 hours of DMSO or XX-650-23 treatment. Representative blots of at least three independent experiments are shown. F) RT-PCR confirmed downregulation of CREB-bound genes identified on RNA-Seq following 12 hours of XX-650-23 treatment of KG-1 cells for all genes shown. Data are graphed as mean ± SEM ( n = 3), * p
    Figure Legend Snippet: Specificity of CREB Inhibition A) The consensus sequence obtained following CREB ChIP-Seq, mapped against the canonical CRE element sequence. B) Relationship between H3K27 acetylation and CREB-driven gene expression. RNA-Seq, CREB ChIP-Seq and H3K27 histone acetylation-Seq were performed on KG-1 cells treated with 5 μM XX-650-23 or 0.1% DMSO. Combined analysis defined the set of genes bound by CREB in AML cells, and alterations in transcription and histone acetylation occurring secondary to XX-650-23 at those loci. The percent of histone acetylation remaining following treatment with XX-650-23 is plotted versus the corresponding change in gene expression for all CREB-bound genes identified by CREB ChIP-Seq. C) Changes in H3K27 acetylation for CREB-bound and -unbound genes were averaged and plotted against base-pair distance to transcriptional start sites (TSS) following XX-650-23 or DMSO treatment. Non-CREB-bound genes show no significant change in H3K27 acetylation following XX-650-23 treatment. D) Heatmap of CREB binding and H3K27 acetylation relative to TSS in DMSO and XX-650-23-treated samples. H3K27 signal intensity, but not CREB binding signal intensity, decreased following XX-650-23. E) Western blot of total and H3K27-specific histone acetylation following XX-650-23 treatment following 6 or 24 hours of DMSO or XX-650-23 treatment. Representative blots of at least three independent experiments are shown. F) RT-PCR confirmed downregulation of CREB-bound genes identified on RNA-Seq following 12 hours of XX-650-23 treatment of KG-1 cells for all genes shown. Data are graphed as mean ± SEM ( n = 3), * p

    Techniques Used: Inhibition, Sequencing, Chromatin Immunoprecipitation, Expressing, RNA Sequencing Assay, Binding Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction

    27) Product Images from "SIRT1 Modulates the Sensitivity of Prostate Cancer Cells to Vesicular Stomatitis Virus Oncolysis"

    Article Title: SIRT1 Modulates the Sensitivity of Prostate Cancer Cells to Vesicular Stomatitis Virus Oncolysis

    Journal: Journal of Virology

    doi: 10.1128/JVI.00626-19

    Inhibition mediated by HDAC1 and HDAC3 induces a marked depletion of S-phase cells and increase in the levels of G 2 /M cells in PC-3 cell cycle arrest. (A) The cell cycle distribution of PC-3 cells treated for 24 h with increasing concentrations of SAHA (0 to 5 μM) was analyzed by propidium iodide (PI) staining by flow cytometry. The percentages of cells in the G 0 /G 1 , S, and G 2 /M phases were evaluated by measuring DNA content. Data represent means ± SD of results from three independent experiments. (B and C) PC-3 cells, pretreated as described for panel A, were subsequently infected with VSVΔM51-GFP (MOI of 10 −2 ). Infectivity was quantified at 24 h postinfection (p.i.) by flow cytometry. (B) Cell death was assessed using annexin V (AnnV)/7AAD staining by flow cytometry. (C) Data represent means ± SD of results from three experiments. (D) PC-3 cells treated with SAHA (5 μM) for 24 h (T0), as well as PC-3 cells pretreated with SAHA and then infected with VSVΔM51-GFP (MOI of 10 −2 ) for a subsequent 24 h, were analyzed for CDKN1A gene expression by qPCR. Gene expression levels were calculated using the threshold cycle (ΔΔ C T ) method. Data represent means ± SD of results from three independent experiments. The same samples were used to obtain total cell extracts and analyzed by immunoblotting for p21 protein levels. β-Actin was used as a loading control. Results are from a representative experiment. F.I., fold increase. (E to G) PC-3 cells were treated with SAHA (5 μM), RESM (5 μM), TBSA (10 μM), or MS-275 (10 μM) for 24 h (T0) or were left untreated (DMSO). (E) The cell cycle distribution was analyzed by PI staining by using flow cytometer. (F) CDKN1A, CDK6, and CCDN1 gene expression levels were evaluated by qPCR analysis. The gene expression levels were calculated using the ΔΔ C T  method. Data are representative of results from three independent experiments. (G) Total cell extracts were analyzed by immunoblotting for p21, p16INK4A, IκB α, SQSTM1, and LC3B. GAPDH was used as a loading control. Acetyl-α-tubulin and acetyl-histone H3 were used as controls for analysis of the specific activity of the different HDAC inhibitors. Results are from a representative experiment.
    Figure Legend Snippet: Inhibition mediated by HDAC1 and HDAC3 induces a marked depletion of S-phase cells and increase in the levels of G 2 /M cells in PC-3 cell cycle arrest. (A) The cell cycle distribution of PC-3 cells treated for 24 h with increasing concentrations of SAHA (0 to 5 μM) was analyzed by propidium iodide (PI) staining by flow cytometry. The percentages of cells in the G 0 /G 1 , S, and G 2 /M phases were evaluated by measuring DNA content. Data represent means ± SD of results from three independent experiments. (B and C) PC-3 cells, pretreated as described for panel A, were subsequently infected with VSVΔM51-GFP (MOI of 10 −2 ). Infectivity was quantified at 24 h postinfection (p.i.) by flow cytometry. (B) Cell death was assessed using annexin V (AnnV)/7AAD staining by flow cytometry. (C) Data represent means ± SD of results from three experiments. (D) PC-3 cells treated with SAHA (5 μM) for 24 h (T0), as well as PC-3 cells pretreated with SAHA and then infected with VSVΔM51-GFP (MOI of 10 −2 ) for a subsequent 24 h, were analyzed for CDKN1A gene expression by qPCR. Gene expression levels were calculated using the threshold cycle (ΔΔ C T ) method. Data represent means ± SD of results from three independent experiments. The same samples were used to obtain total cell extracts and analyzed by immunoblotting for p21 protein levels. β-Actin was used as a loading control. Results are from a representative experiment. F.I., fold increase. (E to G) PC-3 cells were treated with SAHA (5 μM), RESM (5 μM), TBSA (10 μM), or MS-275 (10 μM) for 24 h (T0) or were left untreated (DMSO). (E) The cell cycle distribution was analyzed by PI staining by using flow cytometer. (F) CDKN1A, CDK6, and CCDN1 gene expression levels were evaluated by qPCR analysis. The gene expression levels were calculated using the ΔΔ C T method. Data are representative of results from three independent experiments. (G) Total cell extracts were analyzed by immunoblotting for p21, p16INK4A, IκB α, SQSTM1, and LC3B. GAPDH was used as a loading control. Acetyl-α-tubulin and acetyl-histone H3 were used as controls for analysis of the specific activity of the different HDAC inhibitors. Results are from a representative experiment.

    Techniques Used: Inhibition, Staining, Flow Cytometry, Cytometry, Infection, Expressing, Real-time Polymerase Chain Reaction, Mass Spectrometry, Activity Assay

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    Immunocytochemistry:

    Article Title: WD Repeat-containing Protein 5 (WDR5) Localizes to the Midbody and Regulates Abscission *
    Article Snippet: .. All antibodies used for Western blot analysis (WB), immunocytochemistry (ICC), and immunoprecipitation (IP) are as follows: rabbit anti-ALIX (Santa Cruz Biotechnology sc-271975, WB 1:500); rabbit anti-Aurora B (Abiocode R0117-1, WB 1:2000); mouse anti-centriolin (Santa Cruz Biotechnology sc-365521, WB 1:500); rabbit anti-CEP55 (Santa Cruz Biotechnology sc-134622, WB 1:1000); rabbit anti-CHMP1B (gift from W. I. Sundquist UT592 , WB 1:500); mouse anti-CYK4 (Santa Cruz Biotechnology sc-271110, WB 1:1250); rabbit anti-ECT2 (Santa Cruz Biotechnology sc-1005, WB 1:600); mouse anti-GAPDH (Biochain Institute Y3322GAPDH, WB 1:6000); mouse anti-GFP (Roche Applied Science, catalog no. 11814460001, WB 1:1600); rabbit anti-histone H3 (Cell Signaling Technology catalog no. 2650, WB 1:4000); rabbit anti-KIF4A (Bethyl A301-074A, WB 1:2500); rabbit anti-mDPY-30 (Ma Laboratory previously characterized , WB 0.25 μg/ml); rabbit anti-MKLP1 (Santa Cruz Biotechnology sc-867, WB 1:1000); goat anti-mouse IgG (Thermo Scientific, DyLight 680 catalog no. 35518, WB 1:6000); goat anti-mouse IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 115-545-062, Rhodamine Red-X catalog no. 115-295-062, and cyanine Cy5 catalog no. 115-175-146, ICC 1:200); goat anti-rabbit IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 111-545-144 and Rhodamine Red-X catalog no. 111-295-144, ICC 1:200); rabbit IgG (Invitrogen catalog no. 02-6102, IP 4 μg/ml); goat anti-rabbit IgG (Thermo Scientific, DyLight 680 catalog no. 35568, WB 1:6000); rabbit anti-PRC1 (Santa Cruz Biotechnology sc-8356, WB 1:1000); rabbit anti-RbBP5 (Bethyl A300-107A, WB 1:2000); rabbit anti-TSG101 (Epitomics catalog no. 5347-1, WB 1:2500); mouse anti-α-tubulin (Covance MMS-407R, ICC 1:6000); mouse anti-acetylated-α-tubulin (Santa Cruz Biotechnology sc-23950, ICC 1:1000); rabbit anti-β-tubulin (Santa Cruz Biotechnology sc-9104, WB 1:200); rabbit anti-WDR5 (Bethyl A302-429A, IP 4 μg/ml); and rabbit anti-WDR5 (Bethyl A302-430A, WB 1:3500). .. All shRNA constructs were purchased from Sigma as glycerol stocks (pLKO.1-puro vector).

    Blocking Assay:

    Article Title: MAT2A as Key Regulator and Therapeutic Target in MLLr Leukemogenesis
    Article Snippet: .. After blocking, primary antibodies (Tri-Methyl-Histone H3 (Lys4) from Cell Signaling and Anti-Histone H3 (mono methyl K79) antibody, Anti-Histone H3 (di methyl K79) antibody, Anti-Histone H4 (symmetric di methyl R3) antibody, and Anti-Histone H3 antibody from Abcam), and the secondary antibodies (anti-rabbit IgG, IRDye 680RD, polyclonal and ant-rabbit IgG, IRDye 800CW, polyclonal from Li-Cor Biosciences) were used. .. Analysis was carried out at the Li-Cor Odyssey CLx using the Image Studio Software (both Li-Cor Biosciences) for documentation.

    Immunoprecipitation:

    Article Title: WD Repeat-containing Protein 5 (WDR5) Localizes to the Midbody and Regulates Abscission *
    Article Snippet: .. All antibodies used for Western blot analysis (WB), immunocytochemistry (ICC), and immunoprecipitation (IP) are as follows: rabbit anti-ALIX (Santa Cruz Biotechnology sc-271975, WB 1:500); rabbit anti-Aurora B (Abiocode R0117-1, WB 1:2000); mouse anti-centriolin (Santa Cruz Biotechnology sc-365521, WB 1:500); rabbit anti-CEP55 (Santa Cruz Biotechnology sc-134622, WB 1:1000); rabbit anti-CHMP1B (gift from W. I. Sundquist UT592 , WB 1:500); mouse anti-CYK4 (Santa Cruz Biotechnology sc-271110, WB 1:1250); rabbit anti-ECT2 (Santa Cruz Biotechnology sc-1005, WB 1:600); mouse anti-GAPDH (Biochain Institute Y3322GAPDH, WB 1:6000); mouse anti-GFP (Roche Applied Science, catalog no. 11814460001, WB 1:1600); rabbit anti-histone H3 (Cell Signaling Technology catalog no. 2650, WB 1:4000); rabbit anti-KIF4A (Bethyl A301-074A, WB 1:2500); rabbit anti-mDPY-30 (Ma Laboratory previously characterized , WB 0.25 μg/ml); rabbit anti-MKLP1 (Santa Cruz Biotechnology sc-867, WB 1:1000); goat anti-mouse IgG (Thermo Scientific, DyLight 680 catalog no. 35518, WB 1:6000); goat anti-mouse IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 115-545-062, Rhodamine Red-X catalog no. 115-295-062, and cyanine Cy5 catalog no. 115-175-146, ICC 1:200); goat anti-rabbit IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 111-545-144 and Rhodamine Red-X catalog no. 111-295-144, ICC 1:200); rabbit IgG (Invitrogen catalog no. 02-6102, IP 4 μg/ml); goat anti-rabbit IgG (Thermo Scientific, DyLight 680 catalog no. 35568, WB 1:6000); rabbit anti-PRC1 (Santa Cruz Biotechnology sc-8356, WB 1:1000); rabbit anti-RbBP5 (Bethyl A300-107A, WB 1:2000); rabbit anti-TSG101 (Epitomics catalog no. 5347-1, WB 1:2500); mouse anti-α-tubulin (Covance MMS-407R, ICC 1:6000); mouse anti-acetylated-α-tubulin (Santa Cruz Biotechnology sc-23950, ICC 1:1000); rabbit anti-β-tubulin (Santa Cruz Biotechnology sc-9104, WB 1:200); rabbit anti-WDR5 (Bethyl A302-429A, IP 4 μg/ml); and rabbit anti-WDR5 (Bethyl A302-430A, WB 1:3500). .. All shRNA constructs were purchased from Sigma as glycerol stocks (pLKO.1-puro vector).

    Western Blot:

    Article Title: WD Repeat-containing Protein 5 (WDR5) Localizes to the Midbody and Regulates Abscission *
    Article Snippet: .. All antibodies used for Western blot analysis (WB), immunocytochemistry (ICC), and immunoprecipitation (IP) are as follows: rabbit anti-ALIX (Santa Cruz Biotechnology sc-271975, WB 1:500); rabbit anti-Aurora B (Abiocode R0117-1, WB 1:2000); mouse anti-centriolin (Santa Cruz Biotechnology sc-365521, WB 1:500); rabbit anti-CEP55 (Santa Cruz Biotechnology sc-134622, WB 1:1000); rabbit anti-CHMP1B (gift from W. I. Sundquist UT592 , WB 1:500); mouse anti-CYK4 (Santa Cruz Biotechnology sc-271110, WB 1:1250); rabbit anti-ECT2 (Santa Cruz Biotechnology sc-1005, WB 1:600); mouse anti-GAPDH (Biochain Institute Y3322GAPDH, WB 1:6000); mouse anti-GFP (Roche Applied Science, catalog no. 11814460001, WB 1:1600); rabbit anti-histone H3 (Cell Signaling Technology catalog no. 2650, WB 1:4000); rabbit anti-KIF4A (Bethyl A301-074A, WB 1:2500); rabbit anti-mDPY-30 (Ma Laboratory previously characterized , WB 0.25 μg/ml); rabbit anti-MKLP1 (Santa Cruz Biotechnology sc-867, WB 1:1000); goat anti-mouse IgG (Thermo Scientific, DyLight 680 catalog no. 35518, WB 1:6000); goat anti-mouse IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 115-545-062, Rhodamine Red-X catalog no. 115-295-062, and cyanine Cy5 catalog no. 115-175-146, ICC 1:200); goat anti-rabbit IgG (Jackson ImmunoResearch, Alexa Fluor 488 catalog no. 111-545-144 and Rhodamine Red-X catalog no. 111-295-144, ICC 1:200); rabbit IgG (Invitrogen catalog no. 02-6102, IP 4 μg/ml); goat anti-rabbit IgG (Thermo Scientific, DyLight 680 catalog no. 35568, WB 1:6000); rabbit anti-PRC1 (Santa Cruz Biotechnology sc-8356, WB 1:1000); rabbit anti-RbBP5 (Bethyl A300-107A, WB 1:2000); rabbit anti-TSG101 (Epitomics catalog no. 5347-1, WB 1:2500); mouse anti-α-tubulin (Covance MMS-407R, ICC 1:6000); mouse anti-acetylated-α-tubulin (Santa Cruz Biotechnology sc-23950, ICC 1:1000); rabbit anti-β-tubulin (Santa Cruz Biotechnology sc-9104, WB 1:200); rabbit anti-WDR5 (Bethyl A302-429A, IP 4 μg/ml); and rabbit anti-WDR5 (Bethyl A302-430A, WB 1:3500). .. All shRNA constructs were purchased from Sigma as glycerol stocks (pLKO.1-puro vector).

    IA:

    Article Title: Identification of Mushroom body miniature, a zinc-finger protein implicated in brain development of Drosophila
    Article Snippet: .. Fixation and immunostainings of third-instar larval brains were performed as described in ref. by using the affinity-purified anti-Mbm, mouse anti-fasciclin II (Developmental Studies Hybridoma Bank, Iowa City, IA), and mouse anti-phospho-histone H3 (Cell Signaling Technology, Beverly, MA) antibodies. .. The corresponding Alexa488- and Cy3-conjugated secondary antibodies were purchased from Molecular Probes and Dianova (Hamburg, Germany).

    Staining:

    Article Title: O-Linked N-Acetylglucosamine Cycling Regulates Mitotic Spindle Organization *
    Article Snippet: .. Antibodies All antibodies were used at a 1:1000 dilution for immunoblotting. pT288 AurA (1:200 for confocal staining, 3079), Survivin (2802), pS7 CENPA (2187), CENPA (2186), Histone H3 (9717), and pS10 histone H3 (1:500 for confocal staining, 9706) antibodies were all purchased from Cell Signaling Technologies. .. Hec1 (1:200 for confocal staining, ab3613), β-tubulin (1:1000 for confocal staining, ab6046), pT232 AurB (1:200 for confocal staining, ab61074), PLK1 (1:200 for confocal staining, ab17057), pT210 PLK1 (ab39068), pS10 histone H3 (1:5,000 for confocal staining, ab5176), INCENP (ab36453), Borealin (ab70910), CDC2 (ab18), pY15 CDC2 (ab47594), and AurA (ab13824) antibodies were purchased from Abcam.

    Affinity Purification:

    Article Title: Identification of Mushroom body miniature, a zinc-finger protein implicated in brain development of Drosophila
    Article Snippet: .. Fixation and immunostainings of third-instar larval brains were performed as described in ref. by using the affinity-purified anti-Mbm, mouse anti-fasciclin II (Developmental Studies Hybridoma Bank, Iowa City, IA), and mouse anti-phospho-histone H3 (Cell Signaling Technology, Beverly, MA) antibodies. .. The corresponding Alexa488- and Cy3-conjugated secondary antibodies were purchased from Molecular Probes and Dianova (Hamburg, Germany).

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    Cell Signaling Technology Inc rabbit anti acetylated histone 4
    ChIP analysis of the levels of acetylated histones at the BRCA1 promoter following M344 and cisplatin treatments . A) MCF7 and B) A2780s cells treated with 5.0 μM M344 in combination with 2 μg/ml cisplatin for 24 hrs show reduced amounts of BRCA1 promoter DNA bound to acetylated <t>histone</t> 4 (AcH4). Real-time PCR products were run on a 1.6% agarose gel.
    Rabbit Anti Acetylated Histone 4, supplied by Cell Signaling Technology Inc, 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/result/rabbit anti acetylated histone 4/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc rabbit anti acetyl histone 3 ab
    vPK epigenetically regulates to enhance transcription of BZLF1 . (A and B) CLIX-FZ BL cells were transfected with empty vector (EV) or pFLAG-vPK (A) or with scrambled siRNA or si- BGLF4 (B), treated with doxycycline after 24 h (A) or 8 h (B), and harvested at different times after treatment for immunoblotting with the indicated antibodies. Numbers indicate the relative amounts of FLAG-ZEBRA and vPK after normalizing to β-actin. (C and D) HH514-16 BL cells were transfected with empty vector (EV) or pFLAG-vPK and treated with NaB after 24 h (C) or transfected with scrambled siRNA or si- BGLF4 and treated with NaB after 8 h (D). Cells were harvested at indicated times after treatment and examined by qRT-PCR using primers directed against BZLF1 transcript and using the ΔΔ C T method. (E and F) CLIX-G4 eBL cells were left untreated or were treated with NaB or NaB plus doxycycline (to induce expression of vPK) and harvested after 24 h for chromatin immunoprecipitation with control antibody (IgG) or antibody against RNA polymerase II (E), <t>histone</t> 3 (F), H3K9me3 (F), or H3Ac (F). Relative occupancy of BZLF1 promoter ( Zp ) by RNA polymerase II (E) and modified histone 3 (F) was determined via qPCR using primers targeting the BZLF1 promoter. All experiments were performed between 2 and 3 times. Error bars in panels C to F represent 3 technical replicates from 2 experiments.
    Rabbit Anti Acetyl Histone 3 Ab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc mouse anti acetylated lysine mab
    Ab3180 recognizes a set of proteins in cells treated with NAM. (A) An <t>acetylated</t> peptide used as an antigen for developing Ab3180. The peptide corresponds to the residues 3174–3186 of human Ki-67 (long form, P46013-1). The <t>lysine</t> residue in the middle of the peptide (K3180) has been shown to become strongly acetylated upon treatment of cells with NAM, a pan-sirtuin inhibitor [ 10 ]. (B–D) Immunoblot analysis of cell extracts with Ab3180. (B) Benzonase extracts of mock- or NAM-treated HCT116 were analyzed. Ab3180 was preincubated with 10-time excess peptide (GGQKSAKVLMQNQC or GGQKSAKacVLMQNQC) by weight at room temperature for 2 h before use. (C) Benzonase extracts of mock- or NAM-treated HCT116 (wild type: WT) and its derivative (Ki-67 knock-out: Ki-67-KO) were analyzed. (D) Benzonase extracts of Ki-67-mACl (#2) cell mock-treated, treated with NAM, or treated with NAM plus IAA were analyzed. (B–D) A specific set of proteins (marked with bold letters a - e ) were clearly detected. The absence (C, lanes 3–4) or degradation (D, lane 3) of endogenous Ki-67 was verified by immunoblot with <t>anti-Ki-67</t> antibody. (E) Representative immunofluorescence images of Ki-67-mACl cells mock-treated, treated with NAM, or treated with IAA. Cells were stained for Ab3180 antigens. Ki-67-mAID-mClover (Ki-67-mACl) was detected via the fluorescence of mClover. DNA was counterstained with Hoechst 33342. Bar, 10 μm.
    Mouse Anti Acetylated Lysine Mab, 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
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    Cell Signaling Technology Inc acetyl lys40 α tubulin
    Effect of BBR on HDAC activity and acetylation of histone H3 and <t>α-tubulin.</t> (a) Total cells lysates were incubated with various doses of BBR or 5 μM SAHA in HDAC assay buffer for 3 h. HDAC activity was initiated by adding the HDAC substrate and incubating at 37°C for 1 h. HDAC activity was measured by detecting the OD value at 405 nm. (b) MDA-MB-231 cells were treated with 25 or 50 μM BBR for 24 ~ 72 h or 1 μM SAHA for 24 h. Protein expressions of Ac-histone H3, Ac-α-tubulin and β-actin were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot. (c) MDA-MB-231 cells were treated with 50 μM BBR for 48 h, and nuclear and cytosolic extracts were prepared as described in “Methods”. Protein expressions of Ac-histone H3, histone H3, Ac-α-tubulin, α-tubulin, HDAC1, HDAC2, HDAC6, and GAPDH were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot.
    Acetyl Lys40 α Tubulin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ChIP analysis of the levels of acetylated histones at the BRCA1 promoter following M344 and cisplatin treatments . A) MCF7 and B) A2780s cells treated with 5.0 μM M344 in combination with 2 μg/ml cisplatin for 24 hrs show reduced amounts of BRCA1 promoter DNA bound to acetylated histone 4 (AcH4). Real-time PCR products were run on a 1.6% agarose gel.

    Journal: Cancer Cell International

    Article Title: The effect of the histone deacetylase inhibitor M344 on BRCA1 expression in breast and ovarian cancer cells

    doi: 10.1186/1475-2867-11-29

    Figure Lengend Snippet: ChIP analysis of the levels of acetylated histones at the BRCA1 promoter following M344 and cisplatin treatments . A) MCF7 and B) A2780s cells treated with 5.0 μM M344 in combination with 2 μg/ml cisplatin for 24 hrs show reduced amounts of BRCA1 promoter DNA bound to acetylated histone 4 (AcH4). Real-time PCR products were run on a 1.6% agarose gel. "Input" controls: untreated; "No Ab" controls: incubation with agarose beads in the absence of αAcH4; and "αAcH4": incubation with agarose beads and αAcH4. The experiment was repeated twice with similar results.

    Article Snippet: For all subsequent immunoblotting, antibodies were diluted to the appropriate concentration in 5% milk in TBS-T. Blots were incubated with the following primary antibodies for 1 hr at room temperature or overnight at 4°C: mouse-anti BRCA1 (1:200, D-9, Santa Cruz, Santa Cruz, CA), rabbit-anti acetylated Histone 4 (acetyl H4) (1:1000, Upstate Cell Signaling, Lake Placid, NY), and mouse-anti actin (1:5000, Sigma-Aldrich).

    Techniques: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Incubation

    vPK epigenetically regulates to enhance transcription of BZLF1 . (A and B) CLIX-FZ BL cells were transfected with empty vector (EV) or pFLAG-vPK (A) or with scrambled siRNA or si- BGLF4 (B), treated with doxycycline after 24 h (A) or 8 h (B), and harvested at different times after treatment for immunoblotting with the indicated antibodies. Numbers indicate the relative amounts of FLAG-ZEBRA and vPK after normalizing to β-actin. (C and D) HH514-16 BL cells were transfected with empty vector (EV) or pFLAG-vPK and treated with NaB after 24 h (C) or transfected with scrambled siRNA or si- BGLF4 and treated with NaB after 8 h (D). Cells were harvested at indicated times after treatment and examined by qRT-PCR using primers directed against BZLF1 transcript and using the ΔΔ C T method. (E and F) CLIX-G4 eBL cells were left untreated or were treated with NaB or NaB plus doxycycline (to induce expression of vPK) and harvested after 24 h for chromatin immunoprecipitation with control antibody (IgG) or antibody against RNA polymerase II (E), histone 3 (F), H3K9me3 (F), or H3Ac (F). Relative occupancy of BZLF1 promoter ( Zp ) by RNA polymerase II (E) and modified histone 3 (F) was determined via qPCR using primers targeting the BZLF1 promoter. All experiments were performed between 2 and 3 times. Error bars in panels C to F represent 3 technical replicates from 2 experiments.

    Journal: Journal of Virology

    Article Title: Retrograde Regulation by the Viral Protein Kinase Epigenetically Sustains the Epstein-Barr Virus Latency-to-Lytic Switch To Augment Virus Production

    doi: 10.1128/JVI.00572-19

    Figure Lengend Snippet: vPK epigenetically regulates to enhance transcription of BZLF1 . (A and B) CLIX-FZ BL cells were transfected with empty vector (EV) or pFLAG-vPK (A) or with scrambled siRNA or si- BGLF4 (B), treated with doxycycline after 24 h (A) or 8 h (B), and harvested at different times after treatment for immunoblotting with the indicated antibodies. Numbers indicate the relative amounts of FLAG-ZEBRA and vPK after normalizing to β-actin. (C and D) HH514-16 BL cells were transfected with empty vector (EV) or pFLAG-vPK and treated with NaB after 24 h (C) or transfected with scrambled siRNA or si- BGLF4 and treated with NaB after 8 h (D). Cells were harvested at indicated times after treatment and examined by qRT-PCR using primers directed against BZLF1 transcript and using the ΔΔ C T method. (E and F) CLIX-G4 eBL cells were left untreated or were treated with NaB or NaB plus doxycycline (to induce expression of vPK) and harvested after 24 h for chromatin immunoprecipitation with control antibody (IgG) or antibody against RNA polymerase II (E), histone 3 (F), H3K9me3 (F), or H3Ac (F). Relative occupancy of BZLF1 promoter ( Zp ) by RNA polymerase II (E) and modified histone 3 (F) was determined via qPCR using primers targeting the BZLF1 promoter. All experiments were performed between 2 and 3 times. Error bars in panels C to F represent 3 technical replicates from 2 experiments.

    Article Snippet: Antibodies included mouse anti-FLAG Ab (F3165; Sigma), rabbit anti-BGLF4 (vPK) Ab (AP8057b; Abgent), rabbit anti-KAP1 Ab (A300-274A; Bethyl Laboratories), goat anti-KAP1 Ab (A303-838A; Bethyl Laboratories), rabbit anti-ATM Ab (A300-299A; Bethyl Laboratories), goat anti-ATM Ab (A300-136A; Bethyl Laboratories), rabbit anti-p-S1981-ATM Ab (ab81292; Abcam), rabbit anti-p-S2996-ATM Ab as described before , mouse anti-β-actin Ab (AC-15; Sigma), rabbit anti-p-S824-KAP1 Ab (A300-767A; Bethyl Laboratories), mouse anti-EA-D Ab (MAB8186; EMD), rabbit anti-phospho-histone H2A.X Ab (9718S; Cell Signaling Technology), rabbit anti-histone H2A.X Ab (7631S; Cell Signaling Technology), rabbit anti-histone 3 Ab (AB1791; Abcam), rabbit anti-acetyl-histone 3 Ab (9649S; Cell Signaling Technology), rabbit anti-histone 3 (trimethyl K9) Ab (AB8898; Abcam), mouse anti-RNA polymerase II CTD Ab (AB817; Abcam), human polyclonal serum against EBV small viral capsid antigen (VCA), as previously described , mouse anti-ZEBRA Ab (a gift from Paul Farrell), horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (H+L) (AP308P; EMD Millipore), HRP-conjugated goat anti-rabbit IgG (H+L) (AP307P; EMD Millipore), HRP-conjugated protein A (101023; Thermo Fisher), fluorescein isothiocyanate (FITC)-conjugated donkey anti-rabbit IgG (31568; Thermo Fisher), phycoerythrin (PE)-conjugated goat anti-mouse IgG (sc-3738; Santa Cruz), PE-conjugated donkey anti-rabbit IgG (sc-3745; Santa Cruz), FITC-conjugated goat anti-mouse IgG (F0257; Sigma), and Alexa Fluor 647-conjugated goat anti-rabbit IgG (A-21245; Thermo Fisher).

    Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Expressing, Chromatin Immunoprecipitation, Modification, Real-time Polymerase Chain Reaction

    Ab3180 recognizes a set of proteins in cells treated with NAM. (A) An acetylated peptide used as an antigen for developing Ab3180. The peptide corresponds to the residues 3174–3186 of human Ki-67 (long form, P46013-1). The lysine residue in the middle of the peptide (K3180) has been shown to become strongly acetylated upon treatment of cells with NAM, a pan-sirtuin inhibitor [ 10 ]. (B–D) Immunoblot analysis of cell extracts with Ab3180. (B) Benzonase extracts of mock- or NAM-treated HCT116 were analyzed. Ab3180 was preincubated with 10-time excess peptide (GGQKSAKVLMQNQC or GGQKSAKacVLMQNQC) by weight at room temperature for 2 h before use. (C) Benzonase extracts of mock- or NAM-treated HCT116 (wild type: WT) and its derivative (Ki-67 knock-out: Ki-67-KO) were analyzed. (D) Benzonase extracts of Ki-67-mACl (#2) cell mock-treated, treated with NAM, or treated with NAM plus IAA were analyzed. (B–D) A specific set of proteins (marked with bold letters a - e ) were clearly detected. The absence (C, lanes 3–4) or degradation (D, lane 3) of endogenous Ki-67 was verified by immunoblot with anti-Ki-67 antibody. (E) Representative immunofluorescence images of Ki-67-mACl cells mock-treated, treated with NAM, or treated with IAA. Cells were stained for Ab3180 antigens. Ki-67-mAID-mClover (Ki-67-mACl) was detected via the fluorescence of mClover. DNA was counterstained with Hoechst 33342. Bar, 10 μm.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Generation of an antibody recognizing a set of acetylated proteins, including subunits of BAF complexes

    doi: 10.1016/j.bbrep.2019.100720

    Figure Lengend Snippet: Ab3180 recognizes a set of proteins in cells treated with NAM. (A) An acetylated peptide used as an antigen for developing Ab3180. The peptide corresponds to the residues 3174–3186 of human Ki-67 (long form, P46013-1). The lysine residue in the middle of the peptide (K3180) has been shown to become strongly acetylated upon treatment of cells with NAM, a pan-sirtuin inhibitor [ 10 ]. (B–D) Immunoblot analysis of cell extracts with Ab3180. (B) Benzonase extracts of mock- or NAM-treated HCT116 were analyzed. Ab3180 was preincubated with 10-time excess peptide (GGQKSAKVLMQNQC or GGQKSAKacVLMQNQC) by weight at room temperature for 2 h before use. (C) Benzonase extracts of mock- or NAM-treated HCT116 (wild type: WT) and its derivative (Ki-67 knock-out: Ki-67-KO) were analyzed. (D) Benzonase extracts of Ki-67-mACl (#2) cell mock-treated, treated with NAM, or treated with NAM plus IAA were analyzed. (B–D) A specific set of proteins (marked with bold letters a - e ) were clearly detected. The absence (C, lanes 3–4) or degradation (D, lane 3) of endogenous Ki-67 was verified by immunoblot with anti-Ki-67 antibody. (E) Representative immunofluorescence images of Ki-67-mACl cells mock-treated, treated with NAM, or treated with IAA. Cells were stained for Ab3180 antigens. Ki-67-mAID-mClover (Ki-67-mACl) was detected via the fluorescence of mClover. DNA was counterstained with Hoechst 33342. Bar, 10 μm.

    Article Snippet: Immunoblot analysis of those IP fractions were performed as described above using the following primary antibodies: rabbit anti-ARID1A/BAF250A (1:1000, D2A8U; Cell Signaling), mouse anti-BRG1 (1:1000, sc-17796; Santa Cruz Biotechnology), rabbit anti-SMARCC2/BAF170 (1:1000, D809V; Cell Signaling), rabbit anti-SMARCC1/BAF155 (1:1000, D7F8S; Cell Signaling), and mouse anti-Acetylated-Lysine mAb (1:1000, Ac-K-103; Cell Signaling).

    Techniques: Knock-Out, Immunofluorescence, Staining, Fluorescence

    Effect of BBR on HDAC activity and acetylation of histone H3 and α-tubulin. (a) Total cells lysates were incubated with various doses of BBR or 5 μM SAHA in HDAC assay buffer for 3 h. HDAC activity was initiated by adding the HDAC substrate and incubating at 37°C for 1 h. HDAC activity was measured by detecting the OD value at 405 nm. (b) MDA-MB-231 cells were treated with 25 or 50 μM BBR for 24 ~ 72 h or 1 μM SAHA for 24 h. Protein expressions of Ac-histone H3, Ac-α-tubulin and β-actin were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot. (c) MDA-MB-231 cells were treated with 50 μM BBR for 48 h, and nuclear and cytosolic extracts were prepared as described in “Methods”. Protein expressions of Ac-histone H3, histone H3, Ac-α-tubulin, α-tubulin, HDAC1, HDAC2, HDAC6, and GAPDH were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot.

    Journal: Scientific Reports

    Article Title: A gene expression signature-based approach reveals the mechanisms of action of the Chinese herbal medicine berberine

    doi: 10.1038/srep06394

    Figure Lengend Snippet: Effect of BBR on HDAC activity and acetylation of histone H3 and α-tubulin. (a) Total cells lysates were incubated with various doses of BBR or 5 μM SAHA in HDAC assay buffer for 3 h. HDAC activity was initiated by adding the HDAC substrate and incubating at 37°C for 1 h. HDAC activity was measured by detecting the OD value at 405 nm. (b) MDA-MB-231 cells were treated with 25 or 50 μM BBR for 24 ~ 72 h or 1 μM SAHA for 24 h. Protein expressions of Ac-histone H3, Ac-α-tubulin and β-actin were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot. (c) MDA-MB-231 cells were treated with 50 μM BBR for 48 h, and nuclear and cytosolic extracts were prepared as described in “Methods”. Protein expressions of Ac-histone H3, histone H3, Ac-α-tubulin, α-tubulin, HDAC1, HDAC2, HDAC6, and GAPDH were analyzed by Western blotting. Images of each indicated probe were cropped from the same blot.

    Article Snippet: PARP1 (#9542S; 1:1000) and acetyl-Lys40-α-tubulin (#5335; 1:2000) antibodies were purchased from Cell Signaling Technology.

    Techniques: Activity Assay, Incubation, Histone Deacetylase Assay, Multiple Displacement Amplification, Western Blot