anti brachyury d2z3j antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti brachyury d2z3j antibody
    Anti Brachyury D2z3j Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti brachyury d2z3j antibody/product/Cell Signaling Technology Inc
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    anti brachyury d2z3j antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti brachyury d2z3j antibody
    Anti Brachyury D2z3j Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti brachyury d2z3j antibody/product/Cell Signaling Technology Inc
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    brachyury  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc brachyury
    Brachyury, 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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    anti brachyury antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti brachyury antibody
    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of <t>brachyury-T,</t> AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
    Anti Brachyury Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti brachyury antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    anti brachyury antibody - by Bioz Stars, 2023-02
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    1) Product Images from "Proteomics and phosphoproteomics of chordoma biopsies reveal alterations in multiple pathways and aberrant kinases activities"

    Article Title: Proteomics and phosphoproteomics of chordoma biopsies reveal alterations in multiple pathways and aberrant kinases activities

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2022.941046

    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of brachyury-T, AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
    Figure Legend Snippet: KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of brachyury-T, AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.

    Techniques Used: Expressing, Western Blot, Derivative Assay, Mass Spectrometry

    anti t brachyury  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti t brachyury
    Anti T Brachyury, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti t brachyury/product/Cell Signaling Technology Inc
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    anti t brachyury - by Bioz Stars, 2023-02
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    anti brachyury d2z3j rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti brachyury d2z3j rabbit mab
    Anti Brachyury D2z3j Rabbit 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
    https://www.bioz.com/result/anti brachyury d2z3j rabbit mab/product/Cell Signaling Technology Inc
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    anti t  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti t
    Anti T, 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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    brachyury  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc brachyury
    Oligonucleotide synthesis and click reaction. A) Oligonucleotides were custom synthesized with a terminal alkyne either at 3’ or 5’ end of the oligo. Oligonucleotide sequences for both 3’ and 5’ alkyne targeting c-Myc (left panel) and <t>brachyury</t> (right panel). B) Chemical structure of azido-VHL ligand and click reaction conditions. C) Chemical structures of 3’ and 5’ modified oligonucleotides after the click reaction with azido-VHL ligand.
    Brachyury, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/brachyury/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
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    brachyury - by Bioz Stars, 2023-02
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    1) Product Images from "OligoTRAFTACs: A Generalizable Method for Transcription Factor Degradation"

    Article Title: OligoTRAFTACs: A Generalizable Method for Transcription Factor Degradation

    Journal: bioRxiv

    doi: 10.1101/2021.12.20.473482

    Oligonucleotide synthesis and click reaction. A) Oligonucleotides were custom synthesized with a terminal alkyne either at 3’ or 5’ end of the oligo. Oligonucleotide sequences for both 3’ and 5’ alkyne targeting c-Myc (left panel) and brachyury (right panel). B) Chemical structure of azido-VHL ligand and click reaction conditions. C) Chemical structures of 3’ and 5’ modified oligonucleotides after the click reaction with azido-VHL ligand.
    Figure Legend Snippet: Oligonucleotide synthesis and click reaction. A) Oligonucleotides were custom synthesized with a terminal alkyne either at 3’ or 5’ end of the oligo. Oligonucleotide sequences for both 3’ and 5’ alkyne targeting c-Myc (left panel) and brachyury (right panel). B) Chemical structure of azido-VHL ligand and click reaction conditions. C) Chemical structures of 3’ and 5’ modified oligonucleotides after the click reaction with azido-VHL ligand.

    Techniques Used: Oligonucleotide Synthesis, Synthesized, Modification

    EMSA and brachyury-GFP degradation data. A) Click reaction mixtures with or without VHL ligand were loaded on to a 1.2% agarose gel and separated over 1 h at constant 120 mV. B) OT1 through 4 were transfected into HEK293T cells overexpressing brachyury-GFP and lysed after 30 h. Cell lysates were subjected to SDS-PAGE and western blotting followed by probing with antibodies against brachyury and GAPDH.
    Figure Legend Snippet: EMSA and brachyury-GFP degradation data. A) Click reaction mixtures with or without VHL ligand were loaded on to a 1.2% agarose gel and separated over 1 h at constant 120 mV. B) OT1 through 4 were transfected into HEK293T cells overexpressing brachyury-GFP and lysed after 30 h. Cell lysates were subjected to SDS-PAGE and western blotting followed by probing with antibodies against brachyury and GAPDH.

    Techniques Used: Agarose Gel Electrophoresis, Transfection, SDS Page, Western Blot

    A) Brachyury-targeting oligonucleotide used in the oligoTRAFTAC design engaged with brachyury-GFP. Brachyury targeting biotinylated oligonucleotide (BRCH) or its scrambled oligonucleotide (SCRM) incubated with cell lysate and captured by streptavidin agarose beads. B) Two oligoTRAFTACs with 3’ VHL ligand modifications, OT3 (5 PEG unit linker) and OT4 (2 PEG unit linker) were transfected into HEK293T cells and brachyury-GFP levels were analyzed in lysates prepared after 20 h. C) OT3 induced brachyury-GFP degradation as early as 12 h in HEK293T cells. D) Washout experiment after 12 h of OT3 transfection. Cells were incubated continuously for 24 h in transfection medium or OT3 was aspirated after 12 h of transfection and fresh medium added to cells. Washout cells incubated for another 12 h and 24 h prior to harvesting.
    Figure Legend Snippet: A) Brachyury-targeting oligonucleotide used in the oligoTRAFTAC design engaged with brachyury-GFP. Brachyury targeting biotinylated oligonucleotide (BRCH) or its scrambled oligonucleotide (SCRM) incubated with cell lysate and captured by streptavidin agarose beads. B) Two oligoTRAFTACs with 3’ VHL ligand modifications, OT3 (5 PEG unit linker) and OT4 (2 PEG unit linker) were transfected into HEK293T cells and brachyury-GFP levels were analyzed in lysates prepared after 20 h. C) OT3 induced brachyury-GFP degradation as early as 12 h in HEK293T cells. D) Washout experiment after 12 h of OT3 transfection. Cells were incubated continuously for 24 h in transfection medium or OT3 was aspirated after 12 h of transfection and fresh medium added to cells. Washout cells incubated for another 12 h and 24 h prior to harvesting.

    Techniques Used: Incubation, Transfection

    Time course and washout experiments for brachyury targeting OTs. A) OT2 through 4 were transfected at 75 nM into HEK293T cells overexpressing brachyury-GFP and lysed after 12h, 24 h, and 36 h. Lysates were probed for brachyury and GAPDH. B) OT3 was transfected into HEK293T cells and washed out after 6 h and 12 h. Cell lysates were probed with antibodies against brachyury and GAPDH.
    Figure Legend Snippet: Time course and washout experiments for brachyury targeting OTs. A) OT2 through 4 were transfected at 75 nM into HEK293T cells overexpressing brachyury-GFP and lysed after 12h, 24 h, and 36 h. Lysates were probed for brachyury and GAPDH. B) OT3 was transfected into HEK293T cells and washed out after 6 h and 12 h. Cell lysates were probed with antibodies against brachyury and GAPDH.

    Techniques Used: Transfection

    A) HEK293T cells expressing brachyury-GFP were transfected with 75 nM each of OT3 and its scrambled OT6, and lysates were probed for brachyury levels. B) OT3 induced brachyury degradation is VHL-dependent. HEK293T cells were preincubated with and without 10 µM of VHL ligand for 1.5 h prior to OT3 transfection. After 20 h of transfection, cells lysates were prepared and analyzed for brachyury degradation. C) OT3 induces brachyury degradation via the proteasomal pathway: neddylation inhibitor MLN-4924 was preincubated with cells prior to OT3 transfection. After 20 h of transfection of OT3, cells were harvested and analyzed for brachyury levels. D) Brachyury-GFP downregulation was monitored by GFP fluorescence in cells in the presence or absence of MLN-4924.
    Figure Legend Snippet: A) HEK293T cells expressing brachyury-GFP were transfected with 75 nM each of OT3 and its scrambled OT6, and lysates were probed for brachyury levels. B) OT3 induced brachyury degradation is VHL-dependent. HEK293T cells were preincubated with and without 10 µM of VHL ligand for 1.5 h prior to OT3 transfection. After 20 h of transfection, cells lysates were prepared and analyzed for brachyury degradation. C) OT3 induces brachyury degradation via the proteasomal pathway: neddylation inhibitor MLN-4924 was preincubated with cells prior to OT3 transfection. After 20 h of transfection of OT3, cells were harvested and analyzed for brachyury levels. D) Brachyury-GFP downregulation was monitored by GFP fluorescence in cells in the presence or absence of MLN-4924.

    Techniques Used: Expressing, Transfection, Fluorescence

    A) Brachyury-GFP degradation by OTs is sequence dependent. OT3, OT4, and their scrambled OTs (OT5 and OT6) were transfected and lysed after 20 h. Cell lysates were subjected to SDS-PAGE and western blotting. Blots were probed with brachyury and GAPDH antibodies. Quantitation of western blot bands is shown on the right. B) Increasing concentration of OT3 were transfected into UM-Chor1 cells and degradation was evaluated after 24 h.
    Figure Legend Snippet: A) Brachyury-GFP degradation by OTs is sequence dependent. OT3, OT4, and their scrambled OTs (OT5 and OT6) were transfected and lysed after 20 h. Cell lysates were subjected to SDS-PAGE and western blotting. Blots were probed with brachyury and GAPDH antibodies. Quantitation of western blot bands is shown on the right. B) Increasing concentration of OT3 were transfected into UM-Chor1 cells and degradation was evaluated after 24 h.

    Techniques Used: Sequencing, Transfection, SDS Page, Western Blot, Quantitation Assay, Concentration Assay

    A) Increasing concentrations of OT17 were transfected into UM-Chor1 cells and harvested after 24 h, subjected to lysis and analyzed for brachyury downregulation. Brachyury levels were normalized to loading control and presented as a bar graph. B) JHC-7 cells were transfected with OT17 and probe for brachyury levels as explained in (A). C) UM-Chor1 cells were transfected with 60 nM of OT17 and harvested at subsequent different time points as indicated. D) Washout experiment: transfection medium was removed after 12 h of OT17 transfection and UM-Chor1 cells were incubated for another 12 h or 24 h in fresh complete cell culture medium. E) OT17-mediated brachyury degradation is oligonucleotide sequence dependent. UM-Chor1 cells were transfected with OT17 and scrambled OT20, cells were lysed and analyzed as shown. F) OT3- and OT17-induced brachyury ubiquitination. HEK293T cells that overexpress brachyury-GFP were transfected with HA-ubiquitin, followed by the second transfection with OT3 or OT17. After 12 h, cell lysates were subjected to immunoprecipitation using brachyury antibody, and the eluates blotted for the indicated proteins.
    Figure Legend Snippet: A) Increasing concentrations of OT17 were transfected into UM-Chor1 cells and harvested after 24 h, subjected to lysis and analyzed for brachyury downregulation. Brachyury levels were normalized to loading control and presented as a bar graph. B) JHC-7 cells were transfected with OT17 and probe for brachyury levels as explained in (A). C) UM-Chor1 cells were transfected with 60 nM of OT17 and harvested at subsequent different time points as indicated. D) Washout experiment: transfection medium was removed after 12 h of OT17 transfection and UM-Chor1 cells were incubated for another 12 h or 24 h in fresh complete cell culture medium. E) OT17-mediated brachyury degradation is oligonucleotide sequence dependent. UM-Chor1 cells were transfected with OT17 and scrambled OT20, cells were lysed and analyzed as shown. F) OT3- and OT17-induced brachyury ubiquitination. HEK293T cells that overexpress brachyury-GFP were transfected with HA-ubiquitin, followed by the second transfection with OT3 or OT17. After 12 h, cell lysates were subjected to immunoprecipitation using brachyury antibody, and the eluates blotted for the indicated proteins.

    Techniques Used: Transfection, Lysis, Incubation, Cell Culture, Sequencing, Immunoprecipitation

    brachyury  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc brachyury
    Brachyury, 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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    81694s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc 81694s
    KEY RESOURCES TABLE
    81694s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Targeted Degradation of Transcription Factors by TRAFTACs: TRAnscription Factor Targeting Chimeras"

    Article Title: Targeted Degradation of Transcription Factors by TRAFTACs: TRAnscription Factor Targeting Chimeras

    Journal: Cell chemical biology

    doi: 10.1016/j.chembiol.2021.03.011

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Transfection, Expressing, Software

    81694s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc 81694s
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    81694s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/81694s/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    1) Product Images from "Targeted Degradation of Transcription Factors by TRAFTACs: TRAnscription Factor Targeting Chimeras"

    Article Title: Targeted Degradation of Transcription Factors by TRAFTACs: TRAnscription Factor Targeting Chimeras

    Journal: Cell chemical biology

    doi: 10.1016/j.chembiol.2021.03.011

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Transfection, Expressing, Software

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    Cell Signaling Technology Inc anti brachyury d2z3j antibody
    Anti Brachyury D2z3j Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of <t>brachyury-T,</t> AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
    Anti Brachyury Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti t brachyury
    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of <t>brachyury-T,</t> AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
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    Cell Signaling Technology Inc anti brachyury d2z3j rabbit mab
    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of <t>brachyury-T,</t> AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
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    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of <t>brachyury-T,</t> AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.
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    KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of brachyury-T, AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.

    Journal: Frontiers in Oncology

    Article Title: Proteomics and phosphoproteomics of chordoma biopsies reveal alterations in multiple pathways and aberrant kinases activities

    doi: 10.3389/fonc.2022.941046

    Figure Lengend Snippet: KiPNA analysis and the expression validation of specific proteins. (A) Kinase-pathway network analysis (KiPNA) indicating the proteomic and phosphoproteomic signaling network in chordoma. (B) Fourteen and 29 kinases were conjectured to be negatively and positively regulated, respectively. (C) The elevated expression of brachyury-T, AURA, MOK, and CDK9 were verified by western immunoblotting analysis. The samples were derived from six independent patients who were distinct from the patients whose samples were used for mass spectrometry. T, tumor tissues; N, normal tissues. (D) Densitometric quantification of western blot results from (B) and presented relative to β-actin expression. The data represent the mean ± SD of each experiment carried out in triplicate.

    Article Snippet: Briefly, samples were incubated with primary anti-Brachyury antibody (81694, CST, USA, 1:200 dilution) at 37°C for 1 hour and second antibody at 37°C for 30 min. Then, the samples were treated with the DAB Substrate kit (PV-8000, ZSGB-BIO, China).

    Techniques: Expressing, Western Blot, Derivative Assay, Mass Spectrometry

    KEY RESOURCES TABLE

    Journal: Cell chemical biology

    Article Title: Targeted Degradation of Transcription Factors by TRAFTACs: TRAnscription Factor Targeting Chimeras

    doi: 10.1016/j.chembiol.2021.03.011

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Cell lines, other reagents and antibodies were purchased from commercial vendors described in the . table ft1 table-wrap mode="anchored" t5 caption a7 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies ECL rabbit IgG, HRP-linked whole Ab GE Health care Cat#NA934; RRID: AB_772206 ECL mouse IgG, HRP-linked whole Ab GE Health care Cat# NA931; RRID: AB_772210 GAPDH (14C10) Cell Signaling Cat# 2118S; RRID: AB_561053 alpha tubulin: Alexa Fluor 488 Invitrogen Cat#53-4502-82;RRID:AB_1210525 NFkB1 p105/p50 Cell Signaling Cat# 3035S; RRID: AB_330564 NFkB p65 Cell Signaling Cat# 8242S; RRID: AB_10859369 brachyury Cell Signaling Cat# 81694S; RRID: AB_2799983 c-Myc (9E10) Santa Cruz Cat# sc-40; RRID: AB_627268 HaloTag Promega Cat# G921A; RRID: AB_2688011 HA-tag Cell Signaling Cat# 3724S; RRID: AB_1549585 DYKDDDDK Tag Cell Signaling Cat# 14793S; RRID: AB_2572291 Bacterial and virus strains E. coli BL21-RIPL codon plus Agilent Technologies Cat# 230280 NEB 5-alpha New England BioLabs Cat# C2987H Rosetta 2 DE3 Sigma Cat# 71397 Chemicals, peptides, and recombinant proteins Tumor Necrosis Factor-α human recombinant protein Sigma Cat# T0157–10UG dCas9 recombinant protein Crews Lab N/A Ni-NTA agarose QIAGEN Cat# 30250 Cycloheximide Sigma Cat# C104450 RNAiMAX transfection reagent Thermo Fisher Sci Cat# 13778–150 MLN4924 selleckchem Cat# S7109 Janelia Fluor 646 (JF646) HaloTag Ligand Promega Cat# ga1120 Critical commercial assays High-Capacity cDNA Reverse Transcription Kit Applied Biosystem Cat# 4368814 KAPA SYBR FAST qPCR Master Mix Kit Kapa Biosystems Cat# KK4600 Experimental models: cell lines HEK293T ATCC Cat# CRL-3216 Flp-In T-Rex 293 Thermo Fisher Sci Cat# {"type":"entrez-nucleotide","attrs":{"text":"R78007","term_id":"853117","term_text":"R78007"}} R78007 HeLa ATCC Cat# CCL-2 Experimental models: organisms/strains Zebrafish ( Danio rerio ) wildtype strain TL Scott Holley Lab, Yale University N/A Oligonucleotides F_CTdCas9:ACCTGACTATGCTGGAGTG GATAAGAAATACUCAATAGGCTTAGCT ATCGGC Yale School of Medicine N/A R_CTdCas9:ATCAGCGGGTTTAACCGGA AATCUCCAGAGTAGACAGCC Yale School of Medicine N/A F_CTPCDNA5:AGATTTCCGGTTAAACC CGCTGAUCAGCCTCGAC Yale School of Medicine N/A R_CTPCDNA5:AGTATTTCTTATCCACTC CAGCATAGTCAGGUACGTCATAAGGG Yale School of Medicine N/A RNA:DNA chimeric oligos This manuscript Figure S1 pCDNA5-HA-NTdCas9HT7 This manuscript Table S1 pCDNA5-HA-NTdCas9HT7-NLS This manuscript Table S1 Recombinant DNA pET302-6His-dCas9-Halo Deng et al Proc Natl Acad.

    Techniques: Recombinant, Transfection, Expressing, Software