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trf2 d1y5d rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc trf2 d1y5d rabbit mab
    Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and <t>TRF2)</t> proteins ( vi ).
    Trf2 D1y5d Rabbit Mab, 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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    Images

    1) Product Images from "Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk"

    Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms27041841

    Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).
    Figure Legend Snippet: Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).

    Techniques Used: Saline, Immunohistochemistry, Isolation, Expressing, Western Blot, Staining, Control, Binding Assay



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    Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and <t>TRF2)</t> proteins ( vi ).
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    Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and <t>TRF2)</t> proteins ( vi ).
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    Image Search Results


    Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).

    Journal: International Journal of Molecular Sciences

    Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk

    doi: 10.3390/ijms27041841

    Figure Lengend Snippet: Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).

    Article Snippet: 12 , TRF2 (D1Y5D) rabbit mAb , CST , 13136S , 1:1000 , 1:500.

    Techniques: Saline, Immunohistochemistry, Isolation, Expressing, Western Blot, Staining, Control, Binding Assay

    a. Outline of DNaseI footprinting experiment. 32 P-labelled 5’ end highlighted by a red asterisk. Radiolabelled template is incubated with KU and DNAPKcs prior to the addition of Shelterin composed of TRF1, TRF2, RAP1, TIN2, POT1 and TPP1. DNase I digested products are then analysed by denaturing Urea-PAGE b-f. DNaseI footprinting of telomere end-binding complexes. Proteins omitted as indicated. Nucleotides from the 5’ telomeric end indicated.

    Journal: bioRxiv

    Article Title: Chromosome end protection by RAP1-mediated inhibition of DNA-PK

    doi: 10.1101/2024.12.28.630583

    Figure Lengend Snippet: a. Outline of DNaseI footprinting experiment. 32 P-labelled 5’ end highlighted by a red asterisk. Radiolabelled template is incubated with KU and DNAPKcs prior to the addition of Shelterin composed of TRF1, TRF2, RAP1, TIN2, POT1 and TPP1. DNase I digested products are then analysed by denaturing Urea-PAGE b-f. DNaseI footprinting of telomere end-binding complexes. Proteins omitted as indicated. Nucleotides from the 5’ telomeric end indicated.

    Article Snippet: Mouse RAP1 was detected with antibody D9H4 (Rabbit mAb #5433), human RAP1 with A300-306A antibody (Bethyl Laboratories) mouse and human TRF2 with D1Y5D (Cell Signaling 13136), and Donkey anti-Rabbit IgG horseradish peroxidase (NA934V, Cytiva) or Goat anti-Rabbit IgG horseradish peroxidase (31460, Invitrogen) secondary antibodies.

    Techniques: Footprinting, Incubation, Binding Assay

    a. domain organisa-tion of RAP1 and TRF2. b-d. DNaseI footprinting of telomere end-binding complexes. Proteins omitted as indicated. See figure S1 for mutant details. e. protein cross-linking analysis of RAP1 and KU in the presence of DNA. Proteins were mixed with crosslinker and the products of the reaction were separat-ed on a denaturing tris-acetate polyacrylamide gel and analysed by silver staining or immunoblotting as indicated. Arroheads mark the position of crosslinked species containing only KU, or KU and RAP1 as indicated f, g. as in b.

    Journal: bioRxiv

    Article Title: Chromosome end protection by RAP1-mediated inhibition of DNA-PK

    doi: 10.1101/2024.12.28.630583

    Figure Lengend Snippet: a. domain organisa-tion of RAP1 and TRF2. b-d. DNaseI footprinting of telomere end-binding complexes. Proteins omitted as indicated. See figure S1 for mutant details. e. protein cross-linking analysis of RAP1 and KU in the presence of DNA. Proteins were mixed with crosslinker and the products of the reaction were separat-ed on a denaturing tris-acetate polyacrylamide gel and analysed by silver staining or immunoblotting as indicated. Arroheads mark the position of crosslinked species containing only KU, or KU and RAP1 as indicated f, g. as in b.

    Article Snippet: Mouse RAP1 was detected with antibody D9H4 (Rabbit mAb #5433), human RAP1 with A300-306A antibody (Bethyl Laboratories) mouse and human TRF2 with D1Y5D (Cell Signaling 13136), and Donkey anti-Rabbit IgG horseradish peroxidase (NA934V, Cytiva) or Goat anti-Rabbit IgG horseradish peroxidase (31460, Invitrogen) secondary antibodies.

    Techniques: Footprinting, Binding Assay, Mutagenesis, Silver Staining, Western Blot

    a. Outline of the KU pulldown approach. See methods for details b-d. KU-bound proteins from reactions containing KU70/80, DNA-PKcs, XRCC4/LIG4, TRF2, RAP1 and template DNA unless indicated were separated by SDS-PAGE and immunoblotted as indicated. TRF2, RAP1 and LIG4 detected with anti-strep tag antibody, KU70 with anti-FLAG antibody. As shown in (b), associated of TRF2 with KU is mediated by template DNA e. Quantification of the percentage of telomeres involved in chromosome fusions per metaphase upon over-expression of mouse RAP1, RAP1 KR/DE and RAP1 R130E after Crispr- and Cre-mediated deletion of Rap1 and Apollo respectively in ApolloF/F Lig4+/+ MEFs. Data from 3 independent experiments, 10 metaphases per experiment (n = 30 total), with median. Statistics by ordinary One-way ANOVA. ‘ns’ not significant, ****P ≤ 0.0001. See Fig. S8 for further details. f. Quantification of the percentage of telomeres fused per metaphase upon Crispr-mediated deletion of Apollo in p53-/-RPE-1 cells with wildtype RAP1 or the RAP1 KR/DE mutant. Data from 3 independent experiments, 10 metaphases per experiment (n = 30 total), with median. Statistics as in (e). See Fig. S8 for further details g. Model for the direct inhibition of DNA-PK by TRF2/RAP1 at mammalian telomeres. When assembled on telomeric DNA, DNA-PK is bound by the RAP1 myb and BRCT domains, which acts as a circuit breaker that prevents DNA-PK from engaging LIG4.

    Journal: bioRxiv

    Article Title: Chromosome end protection by RAP1-mediated inhibition of DNA-PK

    doi: 10.1101/2024.12.28.630583

    Figure Lengend Snippet: a. Outline of the KU pulldown approach. See methods for details b-d. KU-bound proteins from reactions containing KU70/80, DNA-PKcs, XRCC4/LIG4, TRF2, RAP1 and template DNA unless indicated were separated by SDS-PAGE and immunoblotted as indicated. TRF2, RAP1 and LIG4 detected with anti-strep tag antibody, KU70 with anti-FLAG antibody. As shown in (b), associated of TRF2 with KU is mediated by template DNA e. Quantification of the percentage of telomeres involved in chromosome fusions per metaphase upon over-expression of mouse RAP1, RAP1 KR/DE and RAP1 R130E after Crispr- and Cre-mediated deletion of Rap1 and Apollo respectively in ApolloF/F Lig4+/+ MEFs. Data from 3 independent experiments, 10 metaphases per experiment (n = 30 total), with median. Statistics by ordinary One-way ANOVA. ‘ns’ not significant, ****P ≤ 0.0001. See Fig. S8 for further details. f. Quantification of the percentage of telomeres fused per metaphase upon Crispr-mediated deletion of Apollo in p53-/-RPE-1 cells with wildtype RAP1 or the RAP1 KR/DE mutant. Data from 3 independent experiments, 10 metaphases per experiment (n = 30 total), with median. Statistics as in (e). See Fig. S8 for further details g. Model for the direct inhibition of DNA-PK by TRF2/RAP1 at mammalian telomeres. When assembled on telomeric DNA, DNA-PK is bound by the RAP1 myb and BRCT domains, which acts as a circuit breaker that prevents DNA-PK from engaging LIG4.

    Article Snippet: Mouse RAP1 was detected with antibody D9H4 (Rabbit mAb #5433), human RAP1 with A300-306A antibody (Bethyl Laboratories) mouse and human TRF2 with D1Y5D (Cell Signaling 13136), and Donkey anti-Rabbit IgG horseradish peroxidase (NA934V, Cytiva) or Goat anti-Rabbit IgG horseradish peroxidase (31460, Invitrogen) secondary antibodies.

    Techniques: SDS Page, Strep-tag, Over Expression, CRISPR, Mutagenesis, Inhibition