cgamp  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc cgamp
    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), <t>2′,3′-cGAMP</t> (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), <t>3′,3′-cGAMP</t> (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Cgamp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cgamp/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cgamp - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway"

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    Journal: Nature immunology

    doi: 10.1038/s41590-021-00896-3

    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Figure Legend Snippet: a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).

    Techniques Used: Translocation Assay, Infection, Quantitative RT-PCR, Transfection, Transduction, shRNA, Expressing, Two Tailed Test

    a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).
    Figure Legend Snippet: a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).

    Techniques Used: Western Blot, In Vitro, Transduction, shRNA, Flow Cytometry, Isolation, Fluorescence, Microscopy, Stable Transfection, Expressing, Quantitative RT-PCR, Two Tailed Test

    a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).
    Figure Legend Snippet: a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).

    Techniques Used: shRNA, Injection, Quantitative RT-PCR, Western Blot, Flow Cytometry, Isolation, Two Tailed Test

    cgamp  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 93

    Structured Review

    Cell Signaling Technology Inc cgamp
    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), <t>2′,3′-cGAMP</t> (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), <t>3′,3′-cGAMP</t> (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Cgamp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cgamp/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cgamp - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway"

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    Journal: Nature immunology

    doi: 10.1038/s41590-021-00896-3

    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Figure Legend Snippet: a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).

    Techniques Used: Translocation Assay, Infection, Quantitative RT-PCR, Transfection, Transduction, shRNA, Expressing, Two Tailed Test

    a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).
    Figure Legend Snippet: a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).

    Techniques Used: Western Blot, In Vitro, Transduction, shRNA, Flow Cytometry, Isolation, Fluorescence, Microscopy, Stable Transfection, Expressing, Quantitative RT-PCR, Two Tailed Test

    a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).
    Figure Legend Snippet: a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).

    Techniques Used: shRNA, Injection, Quantitative RT-PCR, Western Blot, Flow Cytometry, Isolation, Two Tailed Test

    cgamp  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cgamp
    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), <t>2′,3′-cGAMP</t> (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), <t>3′,3′-cGAMP</t> (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
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    1) Product Images from "The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway"

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    Journal: Nature immunology

    doi: 10.1038/s41590-021-00896-3

    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Figure Legend Snippet: a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).

    Techniques Used: Translocation Assay, Infection, Quantitative RT-PCR, Transfection, Transduction, shRNA, Expressing, Two Tailed Test

    a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).
    Figure Legend Snippet: a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).

    Techniques Used: Western Blot, In Vitro, Transduction, shRNA, Flow Cytometry, Isolation, Fluorescence, Microscopy, Stable Transfection, Expressing, Quantitative RT-PCR, Two Tailed Test

    a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).
    Figure Legend Snippet: a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).

    Techniques Used: shRNA, Injection, Quantitative RT-PCR, Western Blot, Flow Cytometry, Isolation, Two Tailed Test

    sodium cst sorbent  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc sodium cst sorbent
    Sodium Cst Sorbent, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc cgamp
    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), <t>2′,3′-cGAMP</t> (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), <t>3′,3′-cGAMP</t> (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Cgamp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cgamp/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cgamp - by Bioz Stars, 2023-02
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    Cell Signaling Technology Inc sodium cst sorbent
    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), <t>2′,3′-cGAMP</t> (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), <t>3′,3′-cGAMP</t> (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).
    Sodium Cst Sorbent, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium cst sorbent/product/Cell Signaling Technology Inc
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    a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).

    Journal: Nature immunology

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    doi: 10.1038/s41590-021-00896-3

    Figure Lengend Snippet: a , RNAi screen of 1,001 tumor suppressor genes in HUVEC, represented as ranked mean Z-score for poly (dA:dT)-induced IRF3 nuclear translocation (0.5 μg/ml) for 3 h. b , c , IRF3 nuclear translocation in ( b ) HUVEC and ( c ) L929-mRuby-hIRF3 stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) or infected with hCMV (MOI=5) ( b ) for 3 h. d - f , qRT-PCR of ( d ) IFNB1 , ( e ) CXCL10 and ( f ) CCL5 in HUVEC transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml), or infected with hCMV (MOI=5) for 4 h. g - i , qRT-PCR of ( g ) Ifnb1 , ( h ) Cxcl10 , and ( i ) Mx2 in L929-mRuby-hIRF3 transfected with indicated siRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. j - l , qRT-PCR of ( j ) IFNB1 , ( k ) CXCL10 and ( l ) CCL5 in THP1-Blue ISG transduced with indicated shRNA and stimulated with poly (dA:dT) (0.5 μg/ml), VACV70 (2 μg/ml), 2′,3′-cGAMP (10 μg/ml) for 4 h. m , n , qRT-PCR of Ifnb1 in ( m ) MCA205 and ( n ) B16F10 transduced with the indicated shRNA and stimulated with 2′,3′-cGAMP (10 μg/ml), 3′,3′-cGAMP (10 μg/ml) or DMXAA (50 μg/ml) for 4 h. mRNA levels were normalized to values of ACTB (human) and Rn18s or Actb (mouse), and percent expression calculated from stimulated control values. Data in ( b-n ) represent the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( b - n ).

    Article Snippet: Poly (dA:dT) (P0883) and MG132 (474787) were obtained from Sigma-Aldrich; VACV70 from IDT; Lyovec, 2′,3′-cGAMP, 3′,3′-cGAMP, DMXAA, c-di-GMP, poly (I:C)(LMW), poly (I:C)(HMW), LPS-EB, and FSL-1 from Invivogen; Jetprime from VWR; Lipofectamine 2000 from Life Technologies; 10× RIPA buffer (9806S) from Cell Signaling Technology; Lactacystin (70890) from Cayman Chemical; CellTiter Blue and CellTiter Glo from Promega; K48-TUBE-Flag (UM607), K63-TUBE-Flag (UM604), PR-619 (SI9619), and 1,10-phenanthroline (SI9619) from Life Sensors; FITC Annexin-V apoptosis detection kit (556547) from BD Biosciences; DAPK inhibitor #1 ((4Z)-4-(3-Pyridylmethylene)-2-styryl-oxazol-5-one) and DAPK inhibitor #2 ((4Z)-2-phenyl-4-(pyridin-3-ylmethylidene)-1,3-oxazol-5(4H)-one) from EMD Millipore (324788) and Interbioscreen (STOCK1N-35684), respectively . hCMV MOLD obtained from M. Raftery (Charite Medical School) was prepared as described .

    Techniques: Translocation Assay, Infection, Quantitative RT-PCR, Transfection, Transduction, shRNA, Expressing, Two Tailed Test

    a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).

    Journal: Nature immunology

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    doi: 10.1038/s41590-021-00896-3

    Figure Lengend Snippet: a , b , ( Left ) Immunoblot and ( right ) in vitro cell growth of ( a ) MCA205 and ( b ) B16F10 transduced with indicated shRNA. c , Flow cytometry of tumor-infiltrating CD8 + T cells and CD103 + CD8α + DCs in MCA205 tumor suspensions isolated from WT or Ifnar1 -KO mice on Day 6 (n=6 per group). d , Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e , qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f , ( Left ) Immunoblot and ( right ) qRT-PCR of Ifnb1 in sh Dapk3 #1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g , Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h , ( Left ) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or ( right ) paclitaxel (100 nM) for 48 h. i , Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1 -KO mice and treated with teniposide or paclitaxel (n=6 for vehicle, n=7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative ( a-d, g-i ) or the mean ( a, b, e, f ) of three independent experiments. Values represented mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, e, f, h, i ).

    Article Snippet: Poly (dA:dT) (P0883) and MG132 (474787) were obtained from Sigma-Aldrich; VACV70 from IDT; Lyovec, 2′,3′-cGAMP, 3′,3′-cGAMP, DMXAA, c-di-GMP, poly (I:C)(LMW), poly (I:C)(HMW), LPS-EB, and FSL-1 from Invivogen; Jetprime from VWR; Lipofectamine 2000 from Life Technologies; 10× RIPA buffer (9806S) from Cell Signaling Technology; Lactacystin (70890) from Cayman Chemical; CellTiter Blue and CellTiter Glo from Promega; K48-TUBE-Flag (UM607), K63-TUBE-Flag (UM604), PR-619 (SI9619), and 1,10-phenanthroline (SI9619) from Life Sensors; FITC Annexin-V apoptosis detection kit (556547) from BD Biosciences; DAPK inhibitor #1 ((4Z)-4-(3-Pyridylmethylene)-2-styryl-oxazol-5-one) and DAPK inhibitor #2 ((4Z)-2-phenyl-4-(pyridin-3-ylmethylidene)-1,3-oxazol-5(4H)-one) from EMD Millipore (324788) and Interbioscreen (STOCK1N-35684), respectively . hCMV MOLD obtained from M. Raftery (Charite Medical School) was prepared as described .

    Techniques: Western Blot, In Vitro, Transduction, shRNA, Flow Cytometry, Isolation, Fluorescence, Microscopy, Stable Transfection, Expressing, Quantitative RT-PCR, Two Tailed Test

    a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).

    Journal: Nature immunology

    Article Title: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway

    doi: 10.1038/s41590-021-00896-3

    Figure Lengend Snippet: a , ( Left ) Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT (n=7 per group) or ( right ) STING gt/gt mice (n=6 per group) treated using intratumoral injection of 3′,3′-cGAMP. cGAMP injections were performed on Day 6 in WT mice (5 μg per mouse), and Days 6 and 9 in STING gt/gt mice (10 μg per mouse). b , c , qRT-PCR of Ifnb1 , Cxcl10 or Il6 in shRNA-transduced B16F10 treated with ( b ) teniposide (10 μM) for 24 h or ( c ) paclitaxel (100 nM) for 48 h. d , e , Immunoblot of shRNA-transduced B16F10 treated with ( d ) teniposide (10 μM) for 12 h or ( e ) paclitaxel (1 μM) for 12 h (p-STAT1, STAT1, DAPK3, and STING) or 72 h (p-p65 and p65). Data from two gels are presented ( d , e ). f-k , Tumor volume of shRNA-transduced B16F10 subcutaneously transplanted into WT mice and treated with indicated chemical and antibody (n=6 per group). l , ( Upper ) Flow cytometry of tumor-infiltrating CD8 + T cells and ( lower ) CD103 + CD8α + DCs in B16F10 tumor suspensions isolated from WT mice on Day 10 (n=4 for vehicle, n=6 for teniposide-shControl and sh Dapk3 #1, n=7 for teniposide-sh Sting1 #1 and paclitaxel). m , Flow cytometry of intracellular pTBK1 and pIRF3 in B16F10-GFP tumor cells (CD45 − GFP + ) and tumor-infiltrating DCs (CD45 + CD11b − CD11c + ) in B16F10-GFP tumor suspensions isolated from WT mice on Day 10. Values represent percentage of each total cell population. Data in ( a, d-m ) are representative of or ( b, c ) are the mean of three independent experiments. Values represent mean ± s.d. * P <0.05, ** P <0.01, and *** P <0.001. Statistical comparisons were conducted using two-tailed t -test ( a-c, f-m ).

    Article Snippet: Poly (dA:dT) (P0883) and MG132 (474787) were obtained from Sigma-Aldrich; VACV70 from IDT; Lyovec, 2′,3′-cGAMP, 3′,3′-cGAMP, DMXAA, c-di-GMP, poly (I:C)(LMW), poly (I:C)(HMW), LPS-EB, and FSL-1 from Invivogen; Jetprime from VWR; Lipofectamine 2000 from Life Technologies; 10× RIPA buffer (9806S) from Cell Signaling Technology; Lactacystin (70890) from Cayman Chemical; CellTiter Blue and CellTiter Glo from Promega; K48-TUBE-Flag (UM607), K63-TUBE-Flag (UM604), PR-619 (SI9619), and 1,10-phenanthroline (SI9619) from Life Sensors; FITC Annexin-V apoptosis detection kit (556547) from BD Biosciences; DAPK inhibitor #1 ((4Z)-4-(3-Pyridylmethylene)-2-styryl-oxazol-5-one) and DAPK inhibitor #2 ((4Z)-2-phenyl-4-(pyridin-3-ylmethylidene)-1,3-oxazol-5(4H)-one) from EMD Millipore (324788) and Interbioscreen (STOCK1N-35684), respectively . hCMV MOLD obtained from M. Raftery (Charite Medical School) was prepared as described .

    Techniques: shRNA, Injection, Quantitative RT-PCR, Western Blot, Flow Cytometry, Isolation, Two Tailed Test