Structured Review

Abcam histone 4
ACY-738 and ACY-775 are potent and selective histone deacetylase 6 inhibitors. ( a ) N2a cells were incubated with 1 μM ACY-738, ACY-775, or trichostatin A (TSA) as a positive control. By Western blot the acetylation of α-tubulin (acet α-tub) was assessed, as well as the acetylation of histone 3 (acetH3). Total α-tubulin levels, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and <t>histone</t> 4 (H4) were used as a reference for equal loading. ( b ) The ratio of acetylated α-tubulin to total tubulin levels was calculated and normalized to TSA-treated cells. One-way analysis of variance (ANOVA); *** p
Histone 4, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/histone 4/product/Abcam
Average 93 stars, based on 5 article reviews
Price from $9.99 to $1999.99
histone 4 - by Bioz Stars, 2020-09
93/100 stars

Images

1) Product Images from "Development of Improved HDAC6 Inhibitors as Pharmacological Therapy for Axonal Charcot–Marie–Tooth Disease"

Article Title: Development of Improved HDAC6 Inhibitors as Pharmacological Therapy for Axonal Charcot–Marie–Tooth Disease

Journal: Neurotherapeutics

doi: 10.1007/s13311-016-0501-z

ACY-738 and ACY-775 are potent and selective histone deacetylase 6 inhibitors. ( a ) N2a cells were incubated with 1 μM ACY-738, ACY-775, or trichostatin A (TSA) as a positive control. By Western blot the acetylation of α-tubulin (acet α-tub) was assessed, as well as the acetylation of histone 3 (acetH3). Total α-tubulin levels, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and histone 4 (H4) were used as a reference for equal loading. ( b ) The ratio of acetylated α-tubulin to total tubulin levels was calculated and normalized to TSA-treated cells. One-way analysis of variance (ANOVA); *** p
Figure Legend Snippet: ACY-738 and ACY-775 are potent and selective histone deacetylase 6 inhibitors. ( a ) N2a cells were incubated with 1 μM ACY-738, ACY-775, or trichostatin A (TSA) as a positive control. By Western blot the acetylation of α-tubulin (acet α-tub) was assessed, as well as the acetylation of histone 3 (acetH3). Total α-tubulin levels, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and histone 4 (H4) were used as a reference for equal loading. ( b ) The ratio of acetylated α-tubulin to total tubulin levels was calculated and normalized to TSA-treated cells. One-way analysis of variance (ANOVA); *** p

Techniques Used: Histone Deacetylase Assay, Incubation, Positive Control, Western Blot

Related Articles

Western Blot:

Article Title: Mitochondrially targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large-scale deletions or point mutations
Article Snippet: .. Western blotting analysis was performed as previously (Minczuk et al , ), and membranes were probed with the following primary antibodies: anti-FLAG and anti-HA as above, mouse anti-OXPHOS cocktail (Mitosciences, MS601, 1:300), mouse anti-TOM22 (Abcam, ab10436, 1:5000), mouse anti-β-actin (Sigma, A2228, 1:100,000), rabbit anti-SSB1 (kindly donated by Prof. D. Kang, 1:4000), rabbit anti-histone H4 (Abcam, ab10158; 1:5000). .. Secondary antibodies used were HRP-conjugated goat antibodies to rabbit (Promega, W401B; 1:2000), mouse (Promega, W402B, 1:2000) and rat (Santa Cruz, SC2065, 1:1000).

Article Title: Cross talk between microRNA and epigenetic regulation in adult neurogenesis
Article Snippet: .. Membranes were processed according to the ECL Western Blotting Protocol (GE Healthcare). anti-MeCP2 (ab2828; Abcam), anti-Ezh2 (4905; Cell Signaling Technology), anti–tri-methyl-histone H3 (Lys27, C36B11; 9733; Cell Signaling Technology), and anti-histone H4 (ab10158; Abcam) were used as primary antibodies at a 1:1,000 dilution. .. HRP-labeled secondary antibodies were obtained from Sigma-Aldrich (A0545) and were used at a dilution of 1:5,000.

Incubation:

Article Title: The inhibition of checkpoint activation by telomeres does not involve exclusion of dimethylation of histone H4 lysine 20 (H4K20me2)
Article Snippet: .. Antibodies (2 µg) against H4K20me2 (GeneTex GT282 [RRID: AB_2728656] Lot #41582) or total histone H4 (Abcam ab10158 [RRID: AB_296888] Lot #GR133660-1) were mixed with chromatin and incubated at 4°C while rocking for 4 h. Dynabeads Protein G (50 µl, Life Technologies, Cat. No. REF 10004D) was then added and rocked overnight at 4°C. .. Beads were washed with ChIP lysis buffer, ChIP lysis buffer with 500 mM NaCl, Wash buffer and TE buffer (10 mM Tris, 1 mM EDTA pH 7.5) successively .

other:

Article Title: PP32 and SET/TAF-Iβ proteins regulate the acetylation of newly synthesized histone H4
Article Snippet: Antibodies HA (Sigma clone 12CA5), HAT1 (Abcam ab12164), Histone H3 (Abcam ab7834), Histone H4 (Abcam ab10158), H4ac (Millipore 06-866), H4K5ac (Millipore 07-327), H4K8ac (Millipore 06-760), H4K12ac (Millipore 07-595), H4K16ac (Millipore 07-329), Hsp70 (Cell Signalling 4876), Hsp90 (Santa Cruz sc-7947), NASP (Dr Almouzni), PP32 (Abcam ab5991), SET/TAF-Iβ (Abcam ab1183).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Abcam poly acetyl h4
    HDAC1 acts in tandem with T antigen to antagonize the transcriptional activity of CBP. (A) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen, 1 μg of pcDNA3-HDAC1 and 1 μg of renilla reporter vector. Whole cell extracts were used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter was normalized to 1.0 and the other activities are expressed relative to this. The data represent an average of at least five independent transfections. A schematic representation of the constructs used is shown at the bottom of the figure. (B) HeLa cells were transfected with 2 μg of Gal4-TK-luciferase reporter (lane 1), 2 μg of Gal-CBP (FL) (lane 2), 2 μg of pSG5-T antigen (lane 3) and 1 μg of pcDNA3-HDAC1 (lane 4) and comparative ChIP analysis were performed in parallel with the same number of cells using antibodies that specifically recognize acetylated histone H3 (K 9 and 14), poly-acetylated histone H4 and HDAC1. The immunoprecipitates were analyzed by quantitative PCR as in Figure 3 A. Quantification of the acetyl-H3 and <t>acetyl-H4</t> ChIPs bands (as described in Materials and Methods) is shown at the bottom part of the figure. (C) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 1 μg of renilla reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen and 1 μg of pcDNA3-HDAC1 together with 8 μg of HDAC1(1, 2 and 3) or control siRNA vectors. Total cell extracts were prepared 48 h after transfection and used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter in the presence of the control siRNA vectors was normalized to 1.0 and the other activities are expressed relative to this. The diagrams show the relative protein levels obtained from three independent experiments. (D) HeLa cells were transfected with 8 μg of control siRNA or different HDAC1 siRNA (1, 2 or 3) alone or in combination. Total cells extracts were prepared 80 h after transfection and the levels of endogenous HDAC1 protein were detected by western blot analysis. The decrease of HDAC1 levels reached 90%; however, when the HDAC1 levels were analyzed 48 h after transfection the observed decrease was modest (20–30%) (data not shown). The experiment in (C) was performed 48 h after transfection due to the low activation of TK promoter by Gal-CBP after that time. (E) HeLa cells were transfected as in (A) and the levels of T antigen protein in the presence of control siRNA or HDAC1 siRNA were analyzed by Western blot.
    Poly Acetyl H4, supplied by Abcam, 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/poly acetyl h4/product/Abcam
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    poly acetyl h4 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    96
    Abcam histone h4
    A hypothesized methylation regulatory mechanism of bovine SIX1 . The light green circle shows a specific methylation regulation progress of the bovine SIX1 promoter was regulated by <t>histone</t> H4 and E2F2 during the development. The green arrows indicate the progress of individual development and muscle cells differentiation of Qinchuan cattle. The yellow arrows indicate repression of SIX1 promoter activity by DNA methylation through histone H4 and E2F2 TF during development.
    Histone H4, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histone h4/product/Abcam
    Average 96 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    histone h4 - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    99
    Abcam anti phospho histone ser 10 antibody
    PKC δ phosphorylates <t>Ser-10</t> of histone H3 in vitro. ( A ) and ( B ), Recombinant PKCδ was incubated with ATP and core histone (A) or histone H3 (B). After the in vitro kinase(IVK) assay, the samples were analyzed by immunoblotting (IB) with anti-PKCδ, anti-phospho histone H3 Thr 3 (pH3 T3), anti-phospho histone H3 Ser 10 (pH3 S10), anti-phospho histone H3 Thr 11 (pH3 T11), anti-phospho histone H2B Ser 14 (pH2B S14) or anti-histone H3 antibody. ( C ) Recombinant GST, GST-histone H3(1–15) or GST-histone H3(1–15, S10A) proteins were incubated with recombinant PKCδ. Immunoblottings were probed with anti-phospho histone H3 Ser 10 (pH3 S10) or anti-GST antibodies.
    Anti Phospho Histone Ser 10 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho histone ser 10 antibody/product/Abcam
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    anti phospho histone ser 10 antibody - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    HDAC1 acts in tandem with T antigen to antagonize the transcriptional activity of CBP. (A) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen, 1 μg of pcDNA3-HDAC1 and 1 μg of renilla reporter vector. Whole cell extracts were used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter was normalized to 1.0 and the other activities are expressed relative to this. The data represent an average of at least five independent transfections. A schematic representation of the constructs used is shown at the bottom of the figure. (B) HeLa cells were transfected with 2 μg of Gal4-TK-luciferase reporter (lane 1), 2 μg of Gal-CBP (FL) (lane 2), 2 μg of pSG5-T antigen (lane 3) and 1 μg of pcDNA3-HDAC1 (lane 4) and comparative ChIP analysis were performed in parallel with the same number of cells using antibodies that specifically recognize acetylated histone H3 (K 9 and 14), poly-acetylated histone H4 and HDAC1. The immunoprecipitates were analyzed by quantitative PCR as in Figure 3 A. Quantification of the acetyl-H3 and acetyl-H4 ChIPs bands (as described in Materials and Methods) is shown at the bottom part of the figure. (C) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 1 μg of renilla reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen and 1 μg of pcDNA3-HDAC1 together with 8 μg of HDAC1(1, 2 and 3) or control siRNA vectors. Total cell extracts were prepared 48 h after transfection and used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter in the presence of the control siRNA vectors was normalized to 1.0 and the other activities are expressed relative to this. The diagrams show the relative protein levels obtained from three independent experiments. (D) HeLa cells were transfected with 8 μg of control siRNA or different HDAC1 siRNA (1, 2 or 3) alone or in combination. Total cells extracts were prepared 80 h after transfection and the levels of endogenous HDAC1 protein were detected by western blot analysis. The decrease of HDAC1 levels reached 90%; however, when the HDAC1 levels were analyzed 48 h after transfection the observed decrease was modest (20–30%) (data not shown). The experiment in (C) was performed 48 h after transfection due to the low activation of TK promoter by Gal-CBP after that time. (E) HeLa cells were transfected as in (A) and the levels of T antigen protein in the presence of control siRNA or HDAC1 siRNA were analyzed by Western blot.

    Journal: Nucleic Acids Research

    Article Title: Involvement of chromatin and histone deacetylation in SV40 T antigen transcription regulation

    doi: 10.1093/nar/gkl1113

    Figure Lengend Snippet: HDAC1 acts in tandem with T antigen to antagonize the transcriptional activity of CBP. (A) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen, 1 μg of pcDNA3-HDAC1 and 1 μg of renilla reporter vector. Whole cell extracts were used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter was normalized to 1.0 and the other activities are expressed relative to this. The data represent an average of at least five independent transfections. A schematic representation of the constructs used is shown at the bottom of the figure. (B) HeLa cells were transfected with 2 μg of Gal4-TK-luciferase reporter (lane 1), 2 μg of Gal-CBP (FL) (lane 2), 2 μg of pSG5-T antigen (lane 3) and 1 μg of pcDNA3-HDAC1 (lane 4) and comparative ChIP analysis were performed in parallel with the same number of cells using antibodies that specifically recognize acetylated histone H3 (K 9 and 14), poly-acetylated histone H4 and HDAC1. The immunoprecipitates were analyzed by quantitative PCR as in Figure 3 A. Quantification of the acetyl-H3 and acetyl-H4 ChIPs bands (as described in Materials and Methods) is shown at the bottom part of the figure. (C) HeLa cells were transfected with 2 μg of the Gal4-TK-luciferase reporter, 1 μg of renilla reporter, 2 μg of Gal-CBP (FL), 2 μg of pSG5-T antigen and 1 μg of pcDNA3-HDAC1 together with 8 μg of HDAC1(1, 2 and 3) or control siRNA vectors. Total cell extracts were prepared 48 h after transfection and used in the luciferase-renilla assay. The activity derived from the Gal4-TK-luciferase reporter in the presence of the control siRNA vectors was normalized to 1.0 and the other activities are expressed relative to this. The diagrams show the relative protein levels obtained from three independent experiments. (D) HeLa cells were transfected with 8 μg of control siRNA or different HDAC1 siRNA (1, 2 or 3) alone or in combination. Total cells extracts were prepared 80 h after transfection and the levels of endogenous HDAC1 protein were detected by western blot analysis. The decrease of HDAC1 levels reached 90%; however, when the HDAC1 levels were analyzed 48 h after transfection the observed decrease was modest (20–30%) (data not shown). The experiment in (C) was performed 48 h after transfection due to the low activation of TK promoter by Gal-CBP after that time. (E) HeLa cells were transfected as in (A) and the levels of T antigen protein in the presence of control siRNA or HDAC1 siRNA were analyzed by Western blot.

    Article Snippet: The antibodies used were as follows: rabbit polyclonal antibody against Acetyl H3 (k9, k14) and Poly-acetyl H4 (K5, K8, K12, K16) (Upstate) and HDAC1 (Abcam).

    Techniques: Activity Assay, Transfection, Luciferase, Plasmid Preparation, Derivative Assay, Construct, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Western Blot, Activation Assay

    A hypothesized methylation regulatory mechanism of bovine SIX1 . The light green circle shows a specific methylation regulation progress of the bovine SIX1 promoter was regulated by histone H4 and E2F2 during the development. The green arrows indicate the progress of individual development and muscle cells differentiation of Qinchuan cattle. The yellow arrows indicate repression of SIX1 promoter activity by DNA methylation through histone H4 and E2F2 TF during development.

    Journal: International Journal of Molecular Sciences

    Article Title: Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2

    doi: 10.3390/ijms19010213

    Figure Lengend Snippet: A hypothesized methylation regulatory mechanism of bovine SIX1 . The light green circle shows a specific methylation regulation progress of the bovine SIX1 promoter was regulated by histone H4 and E2F2 during the development. The green arrows indicate the progress of individual development and muscle cells differentiation of Qinchuan cattle. The yellow arrows indicate repression of SIX1 promoter activity by DNA methylation through histone H4 and E2F2 TF during development.

    Article Snippet: The protein-DNA complexes were cross-linked and immunoprecipitated with 4 μg of histone H4 or E2F2 antibodies overnight at 4 °C, and the immunoprecipitated products were collected with Protein A+G coated magnetic beads.

    Techniques: Methylation, Activity Assay, DNA Methylation Assay

    Electrophoretic mobility shift assay (EMSA) and ChIP analyses showing direct binding of histone H4 and E2F2 to the SIX1 promoter in vitro and in vivo . ( a , b ) Lane 1: 5′-biotin labeled probe containing histone H4 and E2F2; Lane 2: histone H4 and E2F2 probes were incubated with nuclear extracts; Lane 3: the presence of histone H4 and E2F2 mutation probes (50×); Lane 4: histone H4 and E2F2 probes and nuclear extracts with 50-fold unlabeled oligonucleotides; Lane 5: histone H4 and E2F2 probes and nuclear extracts with 10 μg of anti-histone H4 or anti-E2F2 antibodies. The arrows mark the primary complex and super-shift brand. ( c , d ) ChIP-PCR products were analyzed with the input and immunoprecipitated products for histone H4 and E2F2. The enrichment of DNA fragments in samples immunoprecipitated with histone H4 ( e ) and E2F2 ( f ) antibodies via ChIP-qPCR. Normal rabbit IgG and an intragenic DNA fragment of SIX1 exon 2 were used as negative controls. “**” p

    Journal: International Journal of Molecular Sciences

    Article Title: Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2

    doi: 10.3390/ijms19010213

    Figure Lengend Snippet: Electrophoretic mobility shift assay (EMSA) and ChIP analyses showing direct binding of histone H4 and E2F2 to the SIX1 promoter in vitro and in vivo . ( a , b ) Lane 1: 5′-biotin labeled probe containing histone H4 and E2F2; Lane 2: histone H4 and E2F2 probes were incubated with nuclear extracts; Lane 3: the presence of histone H4 and E2F2 mutation probes (50×); Lane 4: histone H4 and E2F2 probes and nuclear extracts with 50-fold unlabeled oligonucleotides; Lane 5: histone H4 and E2F2 probes and nuclear extracts with 10 μg of anti-histone H4 or anti-E2F2 antibodies. The arrows mark the primary complex and super-shift brand. ( c , d ) ChIP-PCR products were analyzed with the input and immunoprecipitated products for histone H4 and E2F2. The enrichment of DNA fragments in samples immunoprecipitated with histone H4 ( e ) and E2F2 ( f ) antibodies via ChIP-qPCR. Normal rabbit IgG and an intragenic DNA fragment of SIX1 exon 2 were used as negative controls. “**” p

    Article Snippet: The protein-DNA complexes were cross-linked and immunoprecipitated with 4 μg of histone H4 or E2F2 antibodies overnight at 4 °C, and the immunoprecipitated products were collected with Protein A+G coated magnetic beads.

    Techniques: Electrophoretic Mobility Shift Assay, Chromatin Immunoprecipitation, Binding Assay, In Vitro, In Vivo, Labeling, Incubation, Mutagenesis, Polymerase Chain Reaction, Immunoprecipitation, Real-time Polymerase Chain Reaction

    Functional analysis of the histone H4 and E2F2 binding sites as transcriptional repressors in the core promoter region of the SIX1 gene by DNA methylation. ( a ) Sequence and putative transcription factor-binding sites in the core promoter of the SIX1 gene. The putative transcription factor binding sites are boxed. The red sequences indicate methylation loci and capital letters indicate the core sequence of the transcription factors. ( b ) Analyses of SIX1 promoter methylation through unmethylated and methylated luciferase reporter plasmids pGL3−216/−28. ( c ) Luciferase assays in constructs with site-directed mutagenesis of histone H4 and E2F2 binding sites were carried out with the construct pGL3−216/−28 and pGL3M−216/−28. Sequence graphical fill in the black and white are representative of wild and mutant-type, respectively. ( d ) Histone H4 and E2F2 knockdown and overexpression by the specific siRNA and pcDNA3.1 recombinant plasmids and co-transfected with pGL3M−216/−28 in C2C12 cells. The construction of pGL3−216/−28 and pGL3M−216/−28 denote the unmethylated and methylated luciferase reporter plasmids in vitro , respectively. The NC siRNA and pcDNA 3.1 (+) expression plasmid were used as a negative control. The results are expressed as the means ± SD in arbitrary units based on the firefly/Renilla luciferase activity. “*” p

    Journal: International Journal of Molecular Sciences

    Article Title: Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2

    doi: 10.3390/ijms19010213

    Figure Lengend Snippet: Functional analysis of the histone H4 and E2F2 binding sites as transcriptional repressors in the core promoter region of the SIX1 gene by DNA methylation. ( a ) Sequence and putative transcription factor-binding sites in the core promoter of the SIX1 gene. The putative transcription factor binding sites are boxed. The red sequences indicate methylation loci and capital letters indicate the core sequence of the transcription factors. ( b ) Analyses of SIX1 promoter methylation through unmethylated and methylated luciferase reporter plasmids pGL3−216/−28. ( c ) Luciferase assays in constructs with site-directed mutagenesis of histone H4 and E2F2 binding sites were carried out with the construct pGL3−216/−28 and pGL3M−216/−28. Sequence graphical fill in the black and white are representative of wild and mutant-type, respectively. ( d ) Histone H4 and E2F2 knockdown and overexpression by the specific siRNA and pcDNA3.1 recombinant plasmids and co-transfected with pGL3M−216/−28 in C2C12 cells. The construction of pGL3−216/−28 and pGL3M−216/−28 denote the unmethylated and methylated luciferase reporter plasmids in vitro , respectively. The NC siRNA and pcDNA 3.1 (+) expression plasmid were used as a negative control. The results are expressed as the means ± SD in arbitrary units based on the firefly/Renilla luciferase activity. “*” p

    Article Snippet: The protein-DNA complexes were cross-linked and immunoprecipitated with 4 μg of histone H4 or E2F2 antibodies overnight at 4 °C, and the immunoprecipitated products were collected with Protein A+G coated magnetic beads.

    Techniques: Functional Assay, Binding Assay, DNA Methylation Assay, Sequencing, Methylation, Luciferase, Construct, Mutagenesis, Over Expression, Recombinant, Transfection, In Vitro, Expressing, Plasmid Preparation, Negative Control, Activity Assay

    DNA sequence of the coding region of the human histone H4c gene and chemical structures of 1R-Chl and 6R-Chl . A. Chemical structures of 1R-Chl (top) and 6R-Chl (bottom), which target the DNA sequences 5′-WGGWGW-3′ and 5′-WGWGCW-3′, respectively (where W = A or T). Imidazole rings are shown in bold. B. DNA sequence of the coding region of the human histone H4c gene. Potential binding sites for 1R-Chl and 6R-Chl are in bold. The alkylation sites of 1R-Chl and 6R-Chl as verified by LM-PCR are italic-bold and underlined, respectively.

    Journal: Molecular cancer therapeutics

    Article Title: Small molecules targeting Histone H4 as potential therapeutics for chronic myelogenous leukemia

    doi: 10.1158/1535-7163.MCT-08-0130

    Figure Lengend Snippet: DNA sequence of the coding region of the human histone H4c gene and chemical structures of 1R-Chl and 6R-Chl . A. Chemical structures of 1R-Chl (top) and 6R-Chl (bottom), which target the DNA sequences 5′-WGGWGW-3′ and 5′-WGWGCW-3′, respectively (where W = A or T). Imidazole rings are shown in bold. B. DNA sequence of the coding region of the human histone H4c gene. Potential binding sites for 1R-Chl and 6R-Chl are in bold. The alkylation sites of 1R-Chl and 6R-Chl as verified by LM-PCR are italic-bold and underlined, respectively.

    Article Snippet: Membranes were then blocked with 5% bovine serum albumin for 1 h at 4°C and probed with histone H4 or GAPDH (Abcam, CA) primary antibodies.

    Techniques: Sequencing, Binding Assay, Polymerase Chain Reaction

    Effect of polyamides on histone H4c and H4k/j transcript expression and Western blot analysis. A. Chart showing polyamide effect on H4c and H4k/j expression; only 1R-Chl and 6R-Chl caused gene down-regulation. B . K562 cells were treated with 1R-Chl and 6R-Chl , at the indicated concentrations for 24 h, and histone H4 and GAPDH protein levels were assayed by Western blot analysis (top left and bottom right, respectively). After incubating with the secondary antibody, the membrane was washed 3 times and soaked in ECL Western Blotting reagents for 30 sec. The membrane was visualized by autoradiography films after 5–30 sec exposure (5 sec for H4 and 30 sec for GAPDH). Analysis of treatment with 1S-Chl and 3R - Chl (top right and bottom left, respectively) did not show significant decreases in histone H4 levels.

    Journal: Molecular cancer therapeutics

    Article Title: Small molecules targeting Histone H4 as potential therapeutics for chronic myelogenous leukemia

    doi: 10.1158/1535-7163.MCT-08-0130

    Figure Lengend Snippet: Effect of polyamides on histone H4c and H4k/j transcript expression and Western blot analysis. A. Chart showing polyamide effect on H4c and H4k/j expression; only 1R-Chl and 6R-Chl caused gene down-regulation. B . K562 cells were treated with 1R-Chl and 6R-Chl , at the indicated concentrations for 24 h, and histone H4 and GAPDH protein levels were assayed by Western blot analysis (top left and bottom right, respectively). After incubating with the secondary antibody, the membrane was washed 3 times and soaked in ECL Western Blotting reagents for 30 sec. The membrane was visualized by autoradiography films after 5–30 sec exposure (5 sec for H4 and 30 sec for GAPDH). Analysis of treatment with 1S-Chl and 3R - Chl (top right and bottom left, respectively) did not show significant decreases in histone H4 levels.

    Article Snippet: Membranes were then blocked with 5% bovine serum albumin for 1 h at 4°C and probed with histone H4 or GAPDH (Abcam, CA) primary antibodies.

    Techniques: Expressing, Western Blot, Size-exclusion Chromatography, Autoradiography

    PKC δ phosphorylates Ser-10 of histone H3 in vitro. ( A ) and ( B ), Recombinant PKCδ was incubated with ATP and core histone (A) or histone H3 (B). After the in vitro kinase(IVK) assay, the samples were analyzed by immunoblotting (IB) with anti-PKCδ, anti-phospho histone H3 Thr 3 (pH3 T3), anti-phospho histone H3 Ser 10 (pH3 S10), anti-phospho histone H3 Thr 11 (pH3 T11), anti-phospho histone H2B Ser 14 (pH2B S14) or anti-histone H3 antibody. ( C ) Recombinant GST, GST-histone H3(1–15) or GST-histone H3(1–15, S10A) proteins were incubated with recombinant PKCδ. Immunoblottings were probed with anti-phospho histone H3 Ser 10 (pH3 S10) or anti-GST antibodies.

    Journal: PLoS ONE

    Article Title: Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase C?

    doi: 10.1371/journal.pone.0044307

    Figure Lengend Snippet: PKC δ phosphorylates Ser-10 of histone H3 in vitro. ( A ) and ( B ), Recombinant PKCδ was incubated with ATP and core histone (A) or histone H3 (B). After the in vitro kinase(IVK) assay, the samples were analyzed by immunoblotting (IB) with anti-PKCδ, anti-phospho histone H3 Thr 3 (pH3 T3), anti-phospho histone H3 Ser 10 (pH3 S10), anti-phospho histone H3 Thr 11 (pH3 T11), anti-phospho histone H2B Ser 14 (pH2B S14) or anti-histone H3 antibody. ( C ) Recombinant GST, GST-histone H3(1–15) or GST-histone H3(1–15, S10A) proteins were incubated with recombinant PKCδ. Immunoblottings were probed with anti-phospho histone H3 Ser 10 (pH3 S10) or anti-GST antibodies.

    Article Snippet: The samples were incubated overnight at 4°C with anti-phospho histone Ser 10 antibody (Abcam), followed by incubation with Alexa546-conjugated anti-rabbit IgG.

    Techniques: In Vitro, Recombinant, Incubation

    Two distinctive biological implications of the phosphorylation of histone H3 on Ser-10. In response to mitogenic stimuli, histone H3 Ser-10 is phosphorylated by mitotic kinase such as Aurora B or VRK1 to promote the mitotic chromatin condensation (upper). On the other hand, in response to death stimuli, activated PKCδ phosphorylate the same position to facilitate the apoptotic chromatin condensation (lower).

    Journal: PLoS ONE

    Article Title: Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase C?

    doi: 10.1371/journal.pone.0044307

    Figure Lengend Snippet: Two distinctive biological implications of the phosphorylation of histone H3 on Ser-10. In response to mitogenic stimuli, histone H3 Ser-10 is phosphorylated by mitotic kinase such as Aurora B or VRK1 to promote the mitotic chromatin condensation (upper). On the other hand, in response to death stimuli, activated PKCδ phosphorylate the same position to facilitate the apoptotic chromatin condensation (lower).

    Article Snippet: The samples were incubated overnight at 4°C with anti-phospho histone Ser 10 antibody (Abcam), followed by incubation with Alexa546-conjugated anti-rabbit IgG.

    Techniques:

    Ectopic expression of PKCδ induces Ser-10 phosphorylation of histone H3. ( A ) HEK 293A cells were transfected with EGFP vector, EGFP-PKCδ catalytic fragment (CF) and EGFP-PKCδ dominant negative form of catalytic fragment (CF DN), respectively.At 24 hours after transfection, cell lysates were subjected to immunoblot analysis with indicated antibodies. ( B ) Immunofluorescence staining of HEK293A cells transfected with PKCδ constructs. HEK293A cells were transfected with EGFP vector, EGFP-PKCδ CF and EGFP-PKCδ CF DN, respectively. Phosphorylation of histone H3 Ser 10 was monitored with specific antibody. ( C ) Kinase activity of PKCδ is required for histone H3 Ser 10 phosphorylation. Kinase-dead PKCδ CF DN was unable to phosphorylate histone H3 on Ser 10. Numbers of positive cells with anti-phospho histone H3 Ser 10 staining were calculated with means±SD value of three independent experiments. **, P

    Journal: PLoS ONE

    Article Title: Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase C?

    doi: 10.1371/journal.pone.0044307

    Figure Lengend Snippet: Ectopic expression of PKCδ induces Ser-10 phosphorylation of histone H3. ( A ) HEK 293A cells were transfected with EGFP vector, EGFP-PKCδ catalytic fragment (CF) and EGFP-PKCδ dominant negative form of catalytic fragment (CF DN), respectively.At 24 hours after transfection, cell lysates were subjected to immunoblot analysis with indicated antibodies. ( B ) Immunofluorescence staining of HEK293A cells transfected with PKCδ constructs. HEK293A cells were transfected with EGFP vector, EGFP-PKCδ CF and EGFP-PKCδ CF DN, respectively. Phosphorylation of histone H3 Ser 10 was monitored with specific antibody. ( C ) Kinase activity of PKCδ is required for histone H3 Ser 10 phosphorylation. Kinase-dead PKCδ CF DN was unable to phosphorylate histone H3 on Ser 10. Numbers of positive cells with anti-phospho histone H3 Ser 10 staining were calculated with means±SD value of three independent experiments. **, P

    Article Snippet: The samples were incubated overnight at 4°C with anti-phospho histone Ser 10 antibody (Abcam), followed by incubation with Alexa546-conjugated anti-rabbit IgG.

    Techniques: Expressing, Transfection, Plasmid Preparation, Dominant Negative Mutation, Immunofluorescence, Staining, Construct, Activity Assay

    Phosphorylation of histone H3 on Ser-10 in apoptotic cells. ( A ) and ( B ) HeLa cells were treated with 1 mM hydroxyurea for 24 hours and then treated with cisplatin (50 µM) for 12 hours, and TUNEL (A), cleaved caspase-3 (B) and phospho-histone H3 Ser 10 staining cells were detected using immunofluorescence microscope. Upper and lower panel images were taken from the same experimental set (A and B).

    Journal: PLoS ONE

    Article Title: Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase C?

    doi: 10.1371/journal.pone.0044307

    Figure Lengend Snippet: Phosphorylation of histone H3 on Ser-10 in apoptotic cells. ( A ) and ( B ) HeLa cells were treated with 1 mM hydroxyurea for 24 hours and then treated with cisplatin (50 µM) for 12 hours, and TUNEL (A), cleaved caspase-3 (B) and phospho-histone H3 Ser 10 staining cells were detected using immunofluorescence microscope. Upper and lower panel images were taken from the same experimental set (A and B).

    Article Snippet: The samples were incubated overnight at 4°C with anti-phospho histone Ser 10 antibody (Abcam), followed by incubation with Alexa546-conjugated anti-rabbit IgG.

    Techniques: TUNEL Assay, Staining, Immunofluorescence, Microscopy

    PKCδ-mediated phosphorylation of histone H3 on Ser-10 contributes to apoptotic chromatin condensation. HEK293A cells were transfected with EGFP vector, EGFP-PKCδ CF and EGFP-PKCδ CF DN, respectively. Chromatin condensation was analyzed with fluorescent microscope after Hoechst staining. The condensed nuclei were indicated with white arrows (middle panel).

    Journal: PLoS ONE

    Article Title: Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase C?

    doi: 10.1371/journal.pone.0044307

    Figure Lengend Snippet: PKCδ-mediated phosphorylation of histone H3 on Ser-10 contributes to apoptotic chromatin condensation. HEK293A cells were transfected with EGFP vector, EGFP-PKCδ CF and EGFP-PKCδ CF DN, respectively. Chromatin condensation was analyzed with fluorescent microscope after Hoechst staining. The condensed nuclei were indicated with white arrows (middle panel).

    Article Snippet: The samples were incubated overnight at 4°C with anti-phospho histone Ser 10 antibody (Abcam), followed by incubation with Alexa546-conjugated anti-rabbit IgG.

    Techniques: Transfection, Plasmid Preparation, Microscopy, Staining