Structured Review

GeneTex histone h4
GO/CDDP triggered direct association of histones H1/H4 and CDDP with LC3. (A) SDS-PAGE analysis of LC3-immunocomplex from the lysate (Lys) and nuclear (Nuc) fractions of cells with or without GO/CDDP treatment. (B) Western blot analysis of proteins (~11 kDa and ~35 kDa) derived from the nuclear LC3-immunocomplex. (C) Immunofluorescence microscopy of cells with or without GO/CDDP treatment. The cells were immunolabeled with primary antibody for histone H1 or H4 and counter stained with DAPI. (D) Percentages of cells positive with total histone H1, phosphorylated histone H1 (p-histone H1), <t>histone</t> H4 and acetylated histone H4 (H4K16ac). (E) Amount of CDDP associated with nuclear LC3 immunocomplex. The nuclear LC3 immunocomplex was analyzed by ICP-MS to quantify the amount of platinum (Pt) in CDDP. The data represent mean ± s.d of at least 3 independent culture experiments.
Histone H4, supplied by GeneTex, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/histone h4/product/GeneTex
Average 92 stars, based on 2 article reviews
Price from $9.99 to $1999.99
histone h4 - by Bioz Stars, 2020-09
92/100 stars

Images

1) Product Images from "Graphene oxide sensitizes cancer cells to chemotherapeutics by inducing early autophagy events, promoting nuclear trafficking and necrosis"

Article Title: Graphene oxide sensitizes cancer cells to chemotherapeutics by inducing early autophagy events, promoting nuclear trafficking and necrosis

Journal: Theranostics

doi: 10.7150/thno.24173

GO/CDDP triggered direct association of histones H1/H4 and CDDP with LC3. (A) SDS-PAGE analysis of LC3-immunocomplex from the lysate (Lys) and nuclear (Nuc) fractions of cells with or without GO/CDDP treatment. (B) Western blot analysis of proteins (~11 kDa and ~35 kDa) derived from the nuclear LC3-immunocomplex. (C) Immunofluorescence microscopy of cells with or without GO/CDDP treatment. The cells were immunolabeled with primary antibody for histone H1 or H4 and counter stained with DAPI. (D) Percentages of cells positive with total histone H1, phosphorylated histone H1 (p-histone H1), histone H4 and acetylated histone H4 (H4K16ac). (E) Amount of CDDP associated with nuclear LC3 immunocomplex. The nuclear LC3 immunocomplex was analyzed by ICP-MS to quantify the amount of platinum (Pt) in CDDP. The data represent mean ± s.d of at least 3 independent culture experiments.
Figure Legend Snippet: GO/CDDP triggered direct association of histones H1/H4 and CDDP with LC3. (A) SDS-PAGE analysis of LC3-immunocomplex from the lysate (Lys) and nuclear (Nuc) fractions of cells with or without GO/CDDP treatment. (B) Western blot analysis of proteins (~11 kDa and ~35 kDa) derived from the nuclear LC3-immunocomplex. (C) Immunofluorescence microscopy of cells with or without GO/CDDP treatment. The cells were immunolabeled with primary antibody for histone H1 or H4 and counter stained with DAPI. (D) Percentages of cells positive with total histone H1, phosphorylated histone H1 (p-histone H1), histone H4 and acetylated histone H4 (H4K16ac). (E) Amount of CDDP associated with nuclear LC3 immunocomplex. The nuclear LC3 immunocomplex was analyzed by ICP-MS to quantify the amount of platinum (Pt) in CDDP. The data represent mean ± s.d of at least 3 independent culture experiments.

Techniques Used: SDS Page, Western Blot, Derivative Assay, Immunofluorescence, Microscopy, Immunolabeling, Staining, Mass Spectrometry

2) Product Images from "Graphene oxide sensitizes cancer cells to chemotherapeutics by inducing early autophagy events, promoting nuclear trafficking and necrosis"

Article Title: Graphene oxide sensitizes cancer cells to chemotherapeutics by inducing early autophagy events, promoting nuclear trafficking and necrosis

Journal: Theranostics

doi: 10.7150/thno.24173

GO/CDDP triggered direct association of histones H1/H4 and CDDP with LC3. (A) SDS-PAGE analysis of LC3-immunocomplex from the lysate (Lys) and nuclear (Nuc) fractions of cells with or without GO/CDDP treatment. (B) Western blot analysis of proteins (~11 kDa and ~35 kDa) derived from the nuclear LC3-immunocomplex. (C) Immunofluorescence microscopy of cells with or without GO/CDDP treatment. The cells were immunolabeled with primary antibody for histone H1 or H4 and counter stained with DAPI. (D) Percentages of cells positive with total histone H1, phosphorylated histone H1 (p-histone H1), histone H4 and acetylated histone H4 (H4K16ac). (E) Amount of CDDP associated with nuclear LC3 immunocomplex. The nuclear LC3 immunocomplex was analyzed by ICP-MS to quantify the amount of platinum (Pt) in CDDP. The data represent mean ± s.d of at least 3 independent culture experiments.
Figure Legend Snippet: GO/CDDP triggered direct association of histones H1/H4 and CDDP with LC3. (A) SDS-PAGE analysis of LC3-immunocomplex from the lysate (Lys) and nuclear (Nuc) fractions of cells with or without GO/CDDP treatment. (B) Western blot analysis of proteins (~11 kDa and ~35 kDa) derived from the nuclear LC3-immunocomplex. (C) Immunofluorescence microscopy of cells with or without GO/CDDP treatment. The cells were immunolabeled with primary antibody for histone H1 or H4 and counter stained with DAPI. (D) Percentages of cells positive with total histone H1, phosphorylated histone H1 (p-histone H1), histone H4 and acetylated histone H4 (H4K16ac). (E) Amount of CDDP associated with nuclear LC3 immunocomplex. The nuclear LC3 immunocomplex was analyzed by ICP-MS to quantify the amount of platinum (Pt) in CDDP. The data represent mean ± s.d of at least 3 independent culture experiments.

Techniques Used: SDS Page, Western Blot, Derivative Assay, Immunofluorescence, Microscopy, Immunolabeling, Staining, Mass Spectrometry

Related Articles

other:

Article Title: Nrf2 is the key to chemotherapy resistance in MCF7 breast cancer cells under hypoxia
Article Snippet: Antibodies, chemicals, siRNA and plasmids Primary antibodies specific for HIF-1α, Nrf2, p-Nrf2, GCLM, MRP-1, GAPDH, and Histone H3 were purchased from Genetex.

Article Title: Impaired coenzyme A synthesis in fission yeast causes defective mitosis, quiescence-exit failure, histone hypoacetylation and fragile DNA
Article Snippet: Antibody specific for histone H3 (GeneTex GTX21791) was used to detect the amount of histone H3.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89
    GeneTex rbap48
    Physical association between HDAC3 and <t>RbAp48.</t> ( A ) 35 S-labelled in vitro translated HDAC1 (lanes 5 and 6) or HDAC3 (lanes 3–4 and 7–10) was subjected to GST pull down analysis using beads harbouring 1 µg GST–RbAp48 fusion protein (lanes 3, 5 and 8), control GST (lanes 4, 6 and 7), GST–E2F1 AD (lane 9) or GST–CREB AD (lane 10), as indicated. After extensive washing, bound proteins were analysed by SDS–PAGE followed by autoradiography. In lanes 1 and 2, 10% of the amount of in vitro translated HDAC3 or HDAC1 used in the pull down reaction was directly loaded. ( B ) SAOS-2 cells were transiently transfected by calcium phosphate co-precipitation with the indicated expression vector and total cell extracts were immunoprecipitated with the indicated antibody (anti-Flag M2 antibody, antibody F; Sigma). Immunoprecipitates were subjected to western blot analysis using the anti-HA antibody. The arrows indicate the position of the two RbAp48 bands. ( C ) Hela nuclear extracts [50 (lanes 1, 2, 6 and 7) or 200 µl (lanes 11–12)] were immunoprecipitated with 1 µg of either an anti-RbAp48 antibody [lane 2, antibody RBBP (Transduction Laboratories); lane 6, antibody N19 (Santa-Cruz); lane 11, antibody 11G10 (Genetex)] or control anti-HA antibody [lanes 1, 7 and 12, antibody 12CA5 (Roche Diagnostics)]. In lanes 4, 8 and 10, 4 µl of HeLa nuclear extracts were directly loaded. In lanes 3, 5 and 9, 1 µg RBBP, N19 and 11G10 antibodies, respectively, were loaded, to monitor the migration of immunoglobulins. Immunoprecipitates were tested for the presence of HDAC1, HDAC2 and HDAC3 by western blotting using an anti-HDAC antibody (Transduction Laboratories). The stars indicate bands due to the immunoglobulin heavy chains from the anti-RbAp48 antibody (lanes 9 and 11) or the anti-HA antibody (lanes 7 and 12). Note that at longer exposures HDAC1 and HDAC2 could be detected in RBBP immunoprecipitates (lane 2). Also, the amount of HDAC3 in N-19 immunoprecipitates (lane 6) is likely to be overestimated due to co-migration with the immunoglobulin heavy chains, which were weakly detected (lane 5).
    Rbap48, supplied by GeneTex, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rbap48/product/GeneTex
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rbap48 - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    93
    GeneTex anti hdac1
    Proposed working model for the role of C/EBPβ-regulated KLF10 in 3T3-L1 adipocyte differentiation. At an early stage of 3T3-L1 adipocyte differentiation, C/EBPβ transactivates KLF10, which interacts with and recruits <t>HDAC1</t> to the promoter of C/EBPα and inhibits its expression. Because C/EBPα can transactivate PPARγ, the expression of PPARγ could also be inhibited by KLF10 indirectly. Thus, C/EBPβ-regulated KLF10 could contribute to the delayed expression of C/EBPα and PPARγ, which may help to facilitate the successful progression of MCE, an early event during 3T3-L1 adipocyte differentiation. In addition, the C/EBPβ-regulated KLF10 pathway may suggest a self-restricted mechanism for C/EBPβ, which could help to maintain the adipogenesis at an appropriate level.
    Anti Hdac1, supplied by GeneTex, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti hdac1/product/GeneTex
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti hdac1 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    90
    GeneTex phosphor histone h2ax
    DDR plays a major role in Dox-triggered cytotoxicity in GCB-DLBCL, but not ABC-DLBCL, cell lines. CHK2 and <t>H2AX</t> phosphorylation (C) and apoptosis (D) after a 20-hour Dox treatment with or without a 1-hour preincubation with the ATM inhibitor (ATMi), KU-55933. Results shown in the bar graphs are mean ± SD and are representative of 2 independent experiments. Two-tailed Student t test was used for pairwise comparison as indicated. * P
    Phosphor Histone H2ax, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphor histone h2ax/product/GeneTex
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphor histone h2ax - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    Physical association between HDAC3 and RbAp48. ( A ) 35 S-labelled in vitro translated HDAC1 (lanes 5 and 6) or HDAC3 (lanes 3–4 and 7–10) was subjected to GST pull down analysis using beads harbouring 1 µg GST–RbAp48 fusion protein (lanes 3, 5 and 8), control GST (lanes 4, 6 and 7), GST–E2F1 AD (lane 9) or GST–CREB AD (lane 10), as indicated. After extensive washing, bound proteins were analysed by SDS–PAGE followed by autoradiography. In lanes 1 and 2, 10% of the amount of in vitro translated HDAC3 or HDAC1 used in the pull down reaction was directly loaded. ( B ) SAOS-2 cells were transiently transfected by calcium phosphate co-precipitation with the indicated expression vector and total cell extracts were immunoprecipitated with the indicated antibody (anti-Flag M2 antibody, antibody F; Sigma). Immunoprecipitates were subjected to western blot analysis using the anti-HA antibody. The arrows indicate the position of the two RbAp48 bands. ( C ) Hela nuclear extracts [50 (lanes 1, 2, 6 and 7) or 200 µl (lanes 11–12)] were immunoprecipitated with 1 µg of either an anti-RbAp48 antibody [lane 2, antibody RBBP (Transduction Laboratories); lane 6, antibody N19 (Santa-Cruz); lane 11, antibody 11G10 (Genetex)] or control anti-HA antibody [lanes 1, 7 and 12, antibody 12CA5 (Roche Diagnostics)]. In lanes 4, 8 and 10, 4 µl of HeLa nuclear extracts were directly loaded. In lanes 3, 5 and 9, 1 µg RBBP, N19 and 11G10 antibodies, respectively, were loaded, to monitor the migration of immunoglobulins. Immunoprecipitates were tested for the presence of HDAC1, HDAC2 and HDAC3 by western blotting using an anti-HDAC antibody (Transduction Laboratories). The stars indicate bands due to the immunoglobulin heavy chains from the anti-RbAp48 antibody (lanes 9 and 11) or the anti-HA antibody (lanes 7 and 12). Note that at longer exposures HDAC1 and HDAC2 could be detected in RBBP immunoprecipitates (lane 2). Also, the amount of HDAC3 in N-19 immunoprecipitates (lane 6) is likely to be overestimated due to co-migration with the immunoglobulin heavy chains, which were weakly detected (lane 5).

    Journal: Nucleic Acids Research

    Article Title: The histone deacetylase HDAC3 targets RbAp48 to the retinoblastoma protein

    doi:

    Figure Lengend Snippet: Physical association between HDAC3 and RbAp48. ( A ) 35 S-labelled in vitro translated HDAC1 (lanes 5 and 6) or HDAC3 (lanes 3–4 and 7–10) was subjected to GST pull down analysis using beads harbouring 1 µg GST–RbAp48 fusion protein (lanes 3, 5 and 8), control GST (lanes 4, 6 and 7), GST–E2F1 AD (lane 9) or GST–CREB AD (lane 10), as indicated. After extensive washing, bound proteins were analysed by SDS–PAGE followed by autoradiography. In lanes 1 and 2, 10% of the amount of in vitro translated HDAC3 or HDAC1 used in the pull down reaction was directly loaded. ( B ) SAOS-2 cells were transiently transfected by calcium phosphate co-precipitation with the indicated expression vector and total cell extracts were immunoprecipitated with the indicated antibody (anti-Flag M2 antibody, antibody F; Sigma). Immunoprecipitates were subjected to western blot analysis using the anti-HA antibody. The arrows indicate the position of the two RbAp48 bands. ( C ) Hela nuclear extracts [50 (lanes 1, 2, 6 and 7) or 200 µl (lanes 11–12)] were immunoprecipitated with 1 µg of either an anti-RbAp48 antibody [lane 2, antibody RBBP (Transduction Laboratories); lane 6, antibody N19 (Santa-Cruz); lane 11, antibody 11G10 (Genetex)] or control anti-HA antibody [lanes 1, 7 and 12, antibody 12CA5 (Roche Diagnostics)]. In lanes 4, 8 and 10, 4 µl of HeLa nuclear extracts were directly loaded. In lanes 3, 5 and 9, 1 µg RBBP, N19 and 11G10 antibodies, respectively, were loaded, to monitor the migration of immunoglobulins. Immunoprecipitates were tested for the presence of HDAC1, HDAC2 and HDAC3 by western blotting using an anti-HDAC antibody (Transduction Laboratories). The stars indicate bands due to the immunoglobulin heavy chains from the anti-RbAp48 antibody (lanes 9 and 11) or the anti-HA antibody (lanes 7 and 12). Note that at longer exposures HDAC1 and HDAC2 could be detected in RBBP immunoprecipitates (lane 2). Also, the amount of HDAC3 in N-19 immunoprecipitates (lane 6) is likely to be overestimated due to co-migration with the immunoglobulin heavy chains, which were weakly detected (lane 5).

    Article Snippet: These data indicate that RbAp48 is required for repression of E2F activity.

    Techniques: In Vitro, SDS Page, Autoradiography, Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot, Transduction, Migration

    Exogenous HDAC3 increases the Rb–RbAp48 interaction. SAOS-2 cells were transfected with 5 µg of the indicated expression vectors [the Rb expression vector encodes the Rb pocket domain (379–928)] by the calcium phosphate co-precipitation method. The amount of promoter in the transfection was kept constant using empty vectors. Twenty-four hours after transfection, total cell lysates were prepared and immunoprecipitated as described (16), using 1 µg of the indicated antibody [anti-Rb, antibody C15G (Santa Cruz Biotechnologies); anti-HA, antibody 12CA5 (Roche Diagnostics)]. Immunoprecipitates were subjected to western blot analysis using the anti-HA antibody (top) or an anti-Rb antibody (antibody XZ55; Pharmingen) (bottom) using standard procedures. The arrows indicate the position of the exogenous proteins. Note that exogenous Rb migrates at ∼60 kDa, because the expression vector used in these experiments expressed a version of Rb deleted for the first 378 amino acids of the molecule.

    Journal: Nucleic Acids Research

    Article Title: The histone deacetylase HDAC3 targets RbAp48 to the retinoblastoma protein

    doi:

    Figure Lengend Snippet: Exogenous HDAC3 increases the Rb–RbAp48 interaction. SAOS-2 cells were transfected with 5 µg of the indicated expression vectors [the Rb expression vector encodes the Rb pocket domain (379–928)] by the calcium phosphate co-precipitation method. The amount of promoter in the transfection was kept constant using empty vectors. Twenty-four hours after transfection, total cell lysates were prepared and immunoprecipitated as described (16), using 1 µg of the indicated antibody [anti-Rb, antibody C15G (Santa Cruz Biotechnologies); anti-HA, antibody 12CA5 (Roche Diagnostics)]. Immunoprecipitates were subjected to western blot analysis using the anti-HA antibody (top) or an anti-Rb antibody (antibody XZ55; Pharmingen) (bottom) using standard procedures. The arrows indicate the position of the exogenous proteins. Note that exogenous Rb migrates at ∼60 kDa, because the expression vector used in these experiments expressed a version of Rb deleted for the first 378 amino acids of the molecule.

    Article Snippet: These data indicate that RbAp48 is required for repression of E2F activity.

    Techniques: Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot

    Model of transcriptional repression by Rb through the recruitment of histone deacetylases. In G 0 or during the beginning of the G 1 phase of the cell cycle Rb protein (or one of its cousins, ‘Pocket protein’ in the figure) binds to E2F sites (15). It recruits a histone deacetylase (‘HDAC’) through a direct (HDAC1 or HDAC2) or indirect (HDAC3, through RBP1) interaction. These three deacetylases share the ability to recruit the histone-binding protein RbAp48, leading to deacetylation of histones present on the promoter.

    Journal: Nucleic Acids Research

    Article Title: The histone deacetylase HDAC3 targets RbAp48 to the retinoblastoma protein

    doi:

    Figure Lengend Snippet: Model of transcriptional repression by Rb through the recruitment of histone deacetylases. In G 0 or during the beginning of the G 1 phase of the cell cycle Rb protein (or one of its cousins, ‘Pocket protein’ in the figure) binds to E2F sites (15). It recruits a histone deacetylase (‘HDAC’) through a direct (HDAC1 or HDAC2) or indirect (HDAC3, through RBP1) interaction. These three deacetylases share the ability to recruit the histone-binding protein RbAp48, leading to deacetylation of histones present on the promoter.

    Article Snippet: These data indicate that RbAp48 is required for repression of E2F activity.

    Techniques: Histone Deacetylase Assay, Binding Assay

    Proposed working model for the role of C/EBPβ-regulated KLF10 in 3T3-L1 adipocyte differentiation. At an early stage of 3T3-L1 adipocyte differentiation, C/EBPβ transactivates KLF10, which interacts with and recruits HDAC1 to the promoter of C/EBPα and inhibits its expression. Because C/EBPα can transactivate PPARγ, the expression of PPARγ could also be inhibited by KLF10 indirectly. Thus, C/EBPβ-regulated KLF10 could contribute to the delayed expression of C/EBPα and PPARγ, which may help to facilitate the successful progression of MCE, an early event during 3T3-L1 adipocyte differentiation. In addition, the C/EBPβ-regulated KLF10 pathway may suggest a self-restricted mechanism for C/EBPβ, which could help to maintain the adipogenesis at an appropriate level.

    Journal: The Journal of Biological Chemistry

    Article Title: Krüppel-like factor 10 (KLF10) is transactivated by the transcription factor C/EBPβ and involved in early 3T3-L1 preadipocyte differentiation

    doi: 10.1074/jbc.RA118.004401

    Figure Lengend Snippet: Proposed working model for the role of C/EBPβ-regulated KLF10 in 3T3-L1 adipocyte differentiation. At an early stage of 3T3-L1 adipocyte differentiation, C/EBPβ transactivates KLF10, which interacts with and recruits HDAC1 to the promoter of C/EBPα and inhibits its expression. Because C/EBPα can transactivate PPARγ, the expression of PPARγ could also be inhibited by KLF10 indirectly. Thus, C/EBPβ-regulated KLF10 could contribute to the delayed expression of C/EBPα and PPARγ, which may help to facilitate the successful progression of MCE, an early event during 3T3-L1 adipocyte differentiation. In addition, the C/EBPβ-regulated KLF10 pathway may suggest a self-restricted mechanism for C/EBPβ, which could help to maintain the adipogenesis at an appropriate level.

    Article Snippet: Equal amounts of protein were separated by SDS-PAGE; transferred to polyvinylidene fluoride membranes (Millipore); immunoblotted with the indicated antibodies: anti-KLF10 (Abcam, ab73537), anti-C/EBPβ (Santa Cruz, sc-150), anti-C/EBPα (Cell Signaling, #2295), anti-PPARγ (Cell Signaling, #2430), anti-HDAC1 (GeneTex, GTX100513), anti-Hsp90 (Santa Cruz, sc-7947), anti-FLAG (Sigma, # F1904), anti-HA (Santa Cruz, sc-805); and then visualized with horseradish peroxidase–coupled secondary antibodies.

    Techniques: Expressing

    Klf10 promotes the recruitment of HDAC1 to C/EBPα promoter, resulting in decreased enrichment of acetylated histone H4 on C/EBPα promoter. A–C , 3T3-L1 preadipocyte was infected with retrovirus expressing GFP or KLF10 and induced to differentiation. At 24 h postinduction, the cells were harvested for Western blotting ( A ), for ChIP–qPCR ( B ), and by using the indicated antibodies ( C ). D–F , 3T3-L1 preadipocyte was transfected with siNC or siKLF10-1 and induced to differentiation. At 24 h postinduction, the cells were harvested for Western blotting ( D ), for ChIP–qPCR ( E ), and by using the indicated antibodies ( F ). For the ChIP–qPCR assays, enrichment of HDAC1 or acetylated histone H4 ( AcH4 ) on the promoter of C/EBPα was analyzed, and the data were normalized to input DNA. All values are represented as means with error bars representing S.D. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Krüppel-like factor 10 (KLF10) is transactivated by the transcription factor C/EBPβ and involved in early 3T3-L1 preadipocyte differentiation

    doi: 10.1074/jbc.RA118.004401

    Figure Lengend Snippet: Klf10 promotes the recruitment of HDAC1 to C/EBPα promoter, resulting in decreased enrichment of acetylated histone H4 on C/EBPα promoter. A–C , 3T3-L1 preadipocyte was infected with retrovirus expressing GFP or KLF10 and induced to differentiation. At 24 h postinduction, the cells were harvested for Western blotting ( A ), for ChIP–qPCR ( B ), and by using the indicated antibodies ( C ). D–F , 3T3-L1 preadipocyte was transfected with siNC or siKLF10-1 and induced to differentiation. At 24 h postinduction, the cells were harvested for Western blotting ( D ), for ChIP–qPCR ( E ), and by using the indicated antibodies ( F ). For the ChIP–qPCR assays, enrichment of HDAC1 or acetylated histone H4 ( AcH4 ) on the promoter of C/EBPα was analyzed, and the data were normalized to input DNA. All values are represented as means with error bars representing S.D. *, p

    Article Snippet: Equal amounts of protein were separated by SDS-PAGE; transferred to polyvinylidene fluoride membranes (Millipore); immunoblotted with the indicated antibodies: anti-KLF10 (Abcam, ab73537), anti-C/EBPβ (Santa Cruz, sc-150), anti-C/EBPα (Cell Signaling, #2295), anti-PPARγ (Cell Signaling, #2430), anti-HDAC1 (GeneTex, GTX100513), anti-Hsp90 (Santa Cruz, sc-7947), anti-FLAG (Sigma, # F1904), anti-HA (Santa Cruz, sc-805); and then visualized with horseradish peroxidase–coupled secondary antibodies.

    Techniques: Infection, Expressing, Western Blot, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Transfection

    DDR plays a major role in Dox-triggered cytotoxicity in GCB-DLBCL, but not ABC-DLBCL, cell lines. CHK2 and H2AX phosphorylation (C) and apoptosis (D) after a 20-hour Dox treatment with or without a 1-hour preincubation with the ATM inhibitor (ATMi), KU-55933. Results shown in the bar graphs are mean ± SD and are representative of 2 independent experiments. Two-tailed Student t test was used for pairwise comparison as indicated. * P

    Journal: Blood

    Article Title: An oxidative stress-based mechanism of doxorubicin cytotoxicity suggests new therapeutic strategies in ABC-DLBCL

    doi: 10.1182/blood-2016-03-705814

    Figure Lengend Snippet: DDR plays a major role in Dox-triggered cytotoxicity in GCB-DLBCL, but not ABC-DLBCL, cell lines. CHK2 and H2AX phosphorylation (C) and apoptosis (D) after a 20-hour Dox treatment with or without a 1-hour preincubation with the ATM inhibitor (ATMi), KU-55933. Results shown in the bar graphs are mean ± SD and are representative of 2 independent experiments. Two-tailed Student t test was used for pairwise comparison as indicated. * P

    Article Snippet: Primary antibodies used for Immunoblot analysis were purchased from Santa Cruz Biotechnology for STAT3 (sc-482), GAPDH (sc-25778), Mcl-1 (sc-819), JunB (sc-8051), and c-Myc (sc-40); from Cell Signaling Technology for PY-STAT3 (#9131), PS-STAT3 (#9123), P-CHK2 (#2661), CHK2 (#2662), P-p53 (#9284), p53 (#2524), phosphor-histone H2AX (#2577), and H2A; from GeneTex for ATM (GTX70103); and from R & D systems for P-ATM (AF1655).

    Techniques: Two Tailed Test