anti-cleaved caspase-3 Search Results


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    Cell Signaling Technology Inc cleaved caspase 3
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) <t>Caspase-3,</t> green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 1 article reviews
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    99
    Cell Signaling Technology Inc anti cleaved caspase 3
    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) <t>Caspase-3,</t> green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P
    Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cleaved caspase 3/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cleaved caspase 3 - by Bioz Stars, 2021-05
    99/100 stars
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    99
    Cell Signaling Technology Inc rabbit anti cleaved caspase 3
    AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.
    Rabbit Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti cleaved caspase 3/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti cleaved caspase 3 - by Bioz Stars, 2021-05
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    N/A
    Caspase 3 p12 is a protein encoded by the CASP3 gene which is approximately 31 6 kDa Caspase 3 p12 is localised to the cytoplasm It is involved in the
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    N/A
    Rabbit polyclonal antibody to Caspase 3 Cleaved Asp175 Isotype Note IgG Host Note Rabbit Conjugation Note Unconjugated Reactivity Note Human Application Note ELISA IHC P
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    N/A
    Rabbit polyclonal antibody against Caspase 3 protein Immunogen region is N terminal Specificity is as follows for the reactive species H S29
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    N/A
    Involved in the activation cascade of caspases responsible for apoptosis execution Cleaves poly ADP ribose polymerase in vitro as well as lamins Overexpression promotes programmed cell death
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    Image Search Results


    Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Concomitant loss of MLKL and Caspase-8, but not loss of RIPK1 kinase activity or combined loss of RIPK3 and Caspase-8, promotes survival of LUBAC-deficient mice a, Representative images of E10.5 ( n =6 embryos/genotype), scale bar: 2 mm, E14.5 ( n =12 Ripk1 K45A Hoil-1 +/- , n =5 Ripk1 K45A Hoil-1 -/- embryos/genotype) and E15.5 embryos ( n =3 embryos/genotype). Scale bar: 5 mm. *: poor yolk sac vascularisation. b, f, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) at E14.5 (b) or E13.5 (f) and quantification. Mean ± s.e.m. and P values from unpaired two-tailed t -tests (b) or one-way ANOVA (f) are shown. Scale bar: 50 µm. c, FADD-IP in MEFs treated for 3 h with zVAD-fmk ± TNF ( n . d, Cell death by PI incorporation in MEFs ± TNF (10 ng/ml) or LT-α. Mean ± s.e.m. ( n =3 independent experiments) and P values (**** P

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Activity Assay, Mouse Assay, Two Tailed Test

    TNFR1 signalling drives cell death and lethality of HOIL-1-deficient mice at mid-gestation. a, d , Quantification of genotypes of animals obtained from inter-crosses of Tnf -/- Hoil-1 +/- (a) and Tnfr1 -/- Hoil-1 +/- (d) mice. *: dead embryos. b, Representative images of embryos quantified in (a) at E10.5 and E15.5, *; poor yolk sac. c, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E10.5 (n= 3 embryos/genotype). e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in Scale bar: 50 µm. f, Representative images of embryos at E16.5 ( n =2 for Tnfr1 -/- Hoil-1 +/- and n =4 for Tnfr1 -/- Hoil-1 -/- ). Fig. 1g.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: TNFR1 signalling drives cell death and lethality of HOIL-1-deficient mice at mid-gestation. a, d , Quantification of genotypes of animals obtained from inter-crosses of Tnf -/- Hoil-1 +/- (a) and Tnfr1 -/- Hoil-1 +/- (d) mice. *: dead embryos. b, Representative images of embryos quantified in (a) at E10.5 and E15.5, *; poor yolk sac. c, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E10.5 (n= 3 embryos/genotype). e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in Scale bar: 50 µm. f, Representative images of embryos at E16.5 ( n =2 for Tnfr1 -/- Hoil-1 +/- and n =4 for Tnfr1 -/- Hoil-1 -/- ). Fig. 1g.

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Mouse Assay, TUNEL Assay, Staining

    HOIL-1-deficient mice die at mid-gestation a, Schematic representation of the Hoil-1 knockout strategy. Solid boxes represent Hoil-1 exons and grey boxes with a star indicate the targeted exons. Boxes with diagonal and horizontal strips represent LoxP and Frt sites, respectively. b, Specificity of gene recombination was assessed by Southern blotting with 5’ and 3’ probes external to the construct in four clones (14B8, 14F6, 20D7 and 21F7). Digest of the DNA with ApaI, followed by hybridisation with the 3’ probe was expected to show a 5700 bp band for the WT allele and a 7700 bp band for the mutant allele. All four clones appeared to have the correct recombination on the 3’ side. Digest of the DNA with SphI and hybridisation with the 5’ probe was expected to show a 4500 bp WT band and a 6200 bp band for the mutated allele. Clones 14B8, 14F6 and 21F7 appeared to be correctly recombined on the 5’ side. Finally, cutting the DNA with ApaI and hybridising with a hygro probe showed a single band in all clones, indicative of a single integration of the construct in all four ES clones. Clones 14B8 and 14F6 were selected for generation of the two Hoil-1 -/- strains. c , PCR analysis of Hoil-1 wild-type, heterozygous and knockout mice. d , Protein levels of HOIL-1, HOIP and SHARPIN in whole embryo lysates ( n = 3 for Hoil-1 +/- and Hoil-1 -/- embryos and n =1 for Hoil-1 +/+ . e , Quantification of genotypes of animals obtained from inter-crossing C20Hoil-1 +/- mice. *indicates dead embryos. f , Representative images of C20Hoil-1 +/- and C20Hoil-1 -/- embryos from E9.5 to E11.5 as quantified in (e). Scale bar: 2 mm. g, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . h , Whole-mount TUNEL staining of embryos at the indicated stages (embryo/genotype n =2 at E10.5, n =8 for Hoil-1 +/- and n =5 for Hoil-1 -/- at E11.5). Scale bar: 2 mm. i, Quantification of genotypes of animals obtained from inter-crossing Hoil-1 fl/wt Tie2-Cre + with Hoil-1 fl/fl Tie2-Cre - mice. *indicates dead embryos. j , Representative images of embryos with conditional deletion of Hoil-1 in Tie2-Cre expressing cells as quantified in (i). Scale bar: 2 mm. *: poorly vascularised yolk sac. Fig. 1c

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: HOIL-1-deficient mice die at mid-gestation a, Schematic representation of the Hoil-1 knockout strategy. Solid boxes represent Hoil-1 exons and grey boxes with a star indicate the targeted exons. Boxes with diagonal and horizontal strips represent LoxP and Frt sites, respectively. b, Specificity of gene recombination was assessed by Southern blotting with 5’ and 3’ probes external to the construct in four clones (14B8, 14F6, 20D7 and 21F7). Digest of the DNA with ApaI, followed by hybridisation with the 3’ probe was expected to show a 5700 bp band for the WT allele and a 7700 bp band for the mutant allele. All four clones appeared to have the correct recombination on the 3’ side. Digest of the DNA with SphI and hybridisation with the 5’ probe was expected to show a 4500 bp WT band and a 6200 bp band for the mutated allele. Clones 14B8, 14F6 and 21F7 appeared to be correctly recombined on the 5’ side. Finally, cutting the DNA with ApaI and hybridising with a hygro probe showed a single band in all clones, indicative of a single integration of the construct in all four ES clones. Clones 14B8 and 14F6 were selected for generation of the two Hoil-1 -/- strains. c , PCR analysis of Hoil-1 wild-type, heterozygous and knockout mice. d , Protein levels of HOIL-1, HOIP and SHARPIN in whole embryo lysates ( n = 3 for Hoil-1 +/- and Hoil-1 -/- embryos and n =1 for Hoil-1 +/+ . e , Quantification of genotypes of animals obtained from inter-crossing C20Hoil-1 +/- mice. *indicates dead embryos. f , Representative images of C20Hoil-1 +/- and C20Hoil-1 -/- embryos from E9.5 to E11.5 as quantified in (e). Scale bar: 2 mm. g, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . h , Whole-mount TUNEL staining of embryos at the indicated stages (embryo/genotype n =2 at E10.5, n =8 for Hoil-1 +/- and n =5 for Hoil-1 -/- at E11.5). Scale bar: 2 mm. i, Quantification of genotypes of animals obtained from inter-crossing Hoil-1 fl/wt Tie2-Cre + with Hoil-1 fl/fl Tie2-Cre - mice. *indicates dead embryos. j , Representative images of embryos with conditional deletion of Hoil-1 in Tie2-Cre expressing cells as quantified in (i). Scale bar: 2 mm. *: poorly vascularised yolk sac. Fig. 1c

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Mouse Assay, Knock-Out, Southern Blot, Construct, Clone Assay, Hybridization, Mutagenesis, Polymerase Chain Reaction, Staining, TUNEL Assay, Expressing

    Individual deletion of mediators of apoptosis or necroptosis does not prevent cell death and lethality at mid-gestation of HOIL-1- or HOIP-deficient embryos. a , Western blot analysis of MLKL expression in the indicated organs derived from control Mlkl -/- mice ( n . b, d, e, f, Representative images of embryos at different stages of gestation (E10.5: n =7 for Ripk3 -/- Hoil-1 +/- and n =5 for Ripk3 -/- Hoil-1 -/- ; E11.5: n =5 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- ; E12.5: n =9 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- (b), E10.5: n =16 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E11.5: n =8 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E12.5: n =10 for Mlkl -/- Hoip +/- and n =5 for Mlkl -/- Hoip -/- (d), E10.5: n =5 for Caspase-8 +/- Hoip +/- and n =4 for Caspase-8 +/- Hoip -/- ; E11.5: n =6 for Caspase-8 +/- Hoip +/- and n =3 for Caspase-8 +/- Hoip -/- ; E12.5: n =3 for Caspase-8 +/- Hoip +/- and n =2 for Caspase-8 +/- Hoip -/- (e), E10.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =4 for Caspase-8 +/- Hoil-1 -/- ; E11.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =5 for Caspase-8 +/- Hoil-1 -/- ; E12.5: n =6 for Caspase-8 +/- Hoil-1 +/- and n =3 for Caspase-8 +/- Hoil-1 -/- (f)). *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation and cell death at E10.5 as detected by PECAM-1 (red) and cleaved (cl.) Caspase-3 staining (green) (top panel) and whole mount TUNEL staining (bottom panel) ( n =4 per genotype). Scale bar: 50 µm.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Individual deletion of mediators of apoptosis or necroptosis does not prevent cell death and lethality at mid-gestation of HOIL-1- or HOIP-deficient embryos. a , Western blot analysis of MLKL expression in the indicated organs derived from control Mlkl -/- mice ( n . b, d, e, f, Representative images of embryos at different stages of gestation (E10.5: n =7 for Ripk3 -/- Hoil-1 +/- and n =5 for Ripk3 -/- Hoil-1 -/- ; E11.5: n =5 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- ; E12.5: n =9 for Ripk3 -/- Hoil-1 +/- and n =2 for Ripk3 -/- Hoil-1 -/- (b), E10.5: n =16 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E11.5: n =8 for Mlkl -/- Hoip +/- and n =6 for Mlkl -/- Hoip -/- ; E12.5: n =10 for Mlkl -/- Hoip +/- and n =5 for Mlkl -/- Hoip -/- (d), E10.5: n =5 for Caspase-8 +/- Hoip +/- and n =4 for Caspase-8 +/- Hoip -/- ; E11.5: n =6 for Caspase-8 +/- Hoip +/- and n =3 for Caspase-8 +/- Hoip -/- ; E12.5: n =3 for Caspase-8 +/- Hoip +/- and n =2 for Caspase-8 +/- Hoip -/- (e), E10.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =4 for Caspase-8 +/- Hoil-1 -/- ; E11.5: n =2 for Caspase-8 +/- Hoil-1 +/- and n =5 for Caspase-8 +/- Hoil-1 -/- ; E12.5: n =6 for Caspase-8 +/- Hoil-1 +/- and n =3 for Caspase-8 +/- Hoil-1 -/- (f)). *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation and cell death at E10.5 as detected by PECAM-1 (red) and cleaved (cl.) Caspase-3 staining (green) (top panel) and whole mount TUNEL staining (bottom panel) ( n =4 per genotype). Scale bar: 50 µm.

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Western Blot, Expressing, Derivative Assay, Mouse Assay, Staining, TUNEL Assay

    Combined deletion of MLKL and Caspase-8 promotes survival of LUBAC-deficient mice. a, Quantification of genotypes of animals obtained from inter-crosses of Mlkl -/- Caspase-8 +/- Hoip +/- with Mlkl -/- Caspase-8 -/- Hoip +/- mice. *: dead embryos. b, Representative images of adult mice as quantified in (a). c , Kaplan-Meier plot of mouse survival ( n =6 for Mlkl -/- Caspase-8 -/- Hoip -/- and n =9 for Mlkl -/- Caspase-8 -/- Hoil-1 -/- mice). d, Representative images of H E staining of the indicated organs ( n =3 mice/genotype). Scale bar: 200 µm. e, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) (top panel) at E13.5 and respective quantifications (bottom panel). Mean ± s.e.m ( n =5 for Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Mlkl -/- Caspase-8 +/- Hoil-1 -/- ) are shown and results were analysed with unpaired two-tailed t -tests comparing Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- embryos. f, Representative images of H E staining of the indicated organs ( n =3 embryos/genotype). Scale bar: 200 µm. g, Epidermal thickness quantification of mice of the indicated genotypes in (f). Mean ± s.e.m values ( n =3 mice/genotype) are shown and results were analysed with unpaired two-tailed t -tests. h, Western blot analysis of lysates from whole E13.5 embryos of the indicated genotypes as well as L929 cells treated or not with TNF plus zVAD-fmk for 2 h as antibody validation ( n .

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Combined deletion of MLKL and Caspase-8 promotes survival of LUBAC-deficient mice. a, Quantification of genotypes of animals obtained from inter-crosses of Mlkl -/- Caspase-8 +/- Hoip +/- with Mlkl -/- Caspase-8 -/- Hoip +/- mice. *: dead embryos. b, Representative images of adult mice as quantified in (a). c , Kaplan-Meier plot of mouse survival ( n =6 for Mlkl -/- Caspase-8 -/- Hoip -/- and n =9 for Mlkl -/- Caspase-8 -/- Hoil-1 -/- mice). d, Representative images of H E staining of the indicated organs ( n =3 mice/genotype). Scale bar: 200 µm. e, Representative images of yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved (cl.) Caspase-3, green) (top panel) at E13.5 and respective quantifications (bottom panel). Mean ± s.e.m ( n =5 for Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Mlkl -/- Caspase-8 +/- Hoil-1 -/- ) are shown and results were analysed with unpaired two-tailed t -tests comparing Mlkl -/- Caspase-8 -/- Hoil-1 +/- and Mlkl -/- Caspase-8 -/- Hoil-1 -/- embryos. f, Representative images of H E staining of the indicated organs ( n =3 embryos/genotype). Scale bar: 200 µm. g, Epidermal thickness quantification of mice of the indicated genotypes in (f). Mean ± s.e.m values ( n =3 mice/genotype) are shown and results were analysed with unpaired two-tailed t -tests. h, Western blot analysis of lysates from whole E13.5 embryos of the indicated genotypes as well as L929 cells treated or not with TNF plus zVAD-fmk for 2 h as antibody validation ( n .

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Mouse Assay, Staining, Two Tailed Test, Western Blot

    Combined deletion of RIPK3 and Caspase-8 prevents cell death but not embryonic lethality at late gestation that is caused by the loss of HOIL-1. a , Quantification of genotypes of animals obtained from inter-crosses of Ripk3 -/- Caspase-8 +/- Hoil-1 +/- with Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (left panel) or Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (right panel). b, Health status of Ripk3 -/- Caspase-8 +/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- embryos at different developmental stages. c , Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . Scale bar: 50 µm. d, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E14.5 (left panel) and respective quantification (right panel). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported. e, f Representative images (e) and quantification (f) of cell death in different organs as detected by TUNEL staining at E13.5 ( n =3 embryos/genotype) and E14.5 ( n =5 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- , n=2 for Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- lung and liver and n =3 Ripk3 -/- Caspase-8 -/- Hoil-1 -/- heart). Scale bar: 50 µm (e). Mean ± s.e.m. values are shown (f). g, Cell death was analysed by PI staining in MEFs stimulated or not with the indicated ligands for 24 h. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. h, Representative images of H E staining on E13.5 whole-embryo paraffin embedded sections ( n =3 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Ripk3 -/- Caspase-8 +/- Hoil-1 -/- ). *: pericardial effusion, arrows; congested vessels H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Representative images of micro-focus CT scan images of whole E13.5 embryos ( n =3 embryos/genotype). *: pericardial effusion Fig. 3f

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Combined deletion of RIPK3 and Caspase-8 prevents cell death but not embryonic lethality at late gestation that is caused by the loss of HOIL-1. a , Quantification of genotypes of animals obtained from inter-crosses of Ripk3 -/- Caspase-8 +/- Hoil-1 +/- with Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (left panel) or Ripk3 -/- Caspase-8 -/- Hoil-1 +/- mice (right panel). b, Health status of Ripk3 -/- Caspase-8 +/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- embryos at different developmental stages. c , Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . Scale bar: 50 µm. d, Cell death as detected by whole-mount TUNEL staining in yolk sacs at E14.5 (left panel) and respective quantification (right panel). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported. e, f Representative images (e) and quantification (f) of cell death in different organs as detected by TUNEL staining at E13.5 ( n =3 embryos/genotype) and E14.5 ( n =5 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- , n=2 for Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- lung and liver and n =3 Ripk3 -/- Caspase-8 -/- Hoil-1 -/- heart). Scale bar: 50 µm (e). Mean ± s.e.m. values are shown (f). g, Cell death was analysed by PI staining in MEFs stimulated or not with the indicated ligands for 24 h. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. h, Representative images of H E staining on E13.5 whole-embryo paraffin embedded sections ( n =3 for Ripk3 -/- Caspase-8 -/- Hoil-1 +/- and Ripk3 -/- Caspase-8 -/- Hoil-1 -/- and n =2 for Ripk3 -/- Caspase-8 +/- Hoil-1 -/- ). *: pericardial effusion, arrows; congested vessels H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Representative images of micro-focus CT scan images of whole E13.5 embryos ( n =3 embryos/genotype). *: pericardial effusion Fig. 3f

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Mouse Assay, Staining, TUNEL Assay, Computed Tomography

    Ablation of the kinase activity of RIPK1 in HOIL-1- or HOIP-deficient embryos prevents cell death and lethality at mid-gestation but not at late gestation. a, b, Quantification of genotypes of animals obtained after inter-crossing Ripk1 K45A Hoil-1 +/- (a) and Ripk1 K45A Hoip +/- (b) mice. *indicates dead embryos. c, Representative images of embryos quantified in (b) *; poor yolk sac vascularisation. Scale bar: 2 mm. d, Whole-mount TUNEL staining of embryos ( n =2 embryos). Scale bar: 2 mm. e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . f, g, Representative images of cell death in different organs (f) and quantification (g) as detected by TUNEL staining at E14.5 ( n =3 embryos/genotype). Scale bar: 50 μm (f). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported (g). h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *; pericardial effusion, n, necrotic area. H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Cell death was analysed by PI staining in MEFs stimulated or not with TNF for 24 h plus the indicated cell death inhibitors. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. Fig. 3b

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: Ablation of the kinase activity of RIPK1 in HOIL-1- or HOIP-deficient embryos prevents cell death and lethality at mid-gestation but not at late gestation. a, b, Quantification of genotypes of animals obtained after inter-crossing Ripk1 K45A Hoil-1 +/- (a) and Ripk1 K45A Hoip +/- (b) mice. *indicates dead embryos. c, Representative images of embryos quantified in (b) *; poor yolk sac vascularisation. Scale bar: 2 mm. d, Whole-mount TUNEL staining of embryos ( n =2 embryos). Scale bar: 2 mm. e, Single staining showing vascularisation (PECAM-1, top panel) and apoptosis (cleaved (cl.) Caspase-3, bottom panel) of yolk sacs. Merged image is shown in . f, g, Representative images of cell death in different organs (f) and quantification (g) as detected by TUNEL staining at E14.5 ( n =3 embryos/genotype). Scale bar: 50 μm (f). Mean ± s.e.m. ( n =3 embryos/genotype) and P values from one-way ANOVA are reported (g). h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *; pericardial effusion, n, necrotic area. H, heart; L, lung; Li, liver. Scale bar: 200 µm. i, Cell death was analysed by PI staining in MEFs stimulated or not with TNF for 24 h plus the indicated cell death inhibitors. Mean ± s.e.m. ( n =3 independent experiments) and P values from two-way ANOVA are reported. Fig. 3b

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Activity Assay, Mouse Assay, TUNEL Assay, Staining

    HOIL-1 deficiency causes embryonic lethality at mid-gestation due to TNFR1-mediated endothelial cell death a, Mendelian frequencies obtained from inter-crossing Hoil-1 +/- mice, *: dead embryos. b, Representative images of embryos from E9.5 to E11.5 quantified in (a), *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation (PECAM-1, red) and cell death (cleaved (cl.) Caspase-3 staining, green) at E10.5 (top panel) ( n =4 yolk-sacs/genotype), and whole-mount TUNEL staining (bottom, panel) ( n =2 yolk-sacs/genotype). Scale bar: 50 µm. d, g, Quantification of branching points (g) and cleaved Caspase-3 positive cells (g). Mean ± s.e.m. values and P values from unpaired two-tailed t -tests are shown. e, Representative images of embryos at E10.5 ( n =14 Hoil-1 fl/wt Tie2-Cre+ and n =7 Hoil-1 fl/fl Tie2-Cre+ embryos, top panel). *: poor yolk sac vascularisation. Scale bar: 2 mm. Yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (middle panel). Scale bar: 50 µm. Yolk sac whole-mount TUNEL staining ( n =6 Hoil-1 fl/wt Tie2-Cre+ and n =2 Hoil-1 fl/fl Tie2-Cre+ yolk-sacs/genotype , bottom panel). f , Representative images of embryos at E15.5 (top panel, n =6 Tnfr1 -/- Hoil-1 -/- and n =19 Tnfr1 -/- Hoil-1 +/- embryos), scale bar: 2 mm, and yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (bottom panel), Scale bar: 50 µm. h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *: pericardial effusion, arrows; congested vessels. H, heart; L, lung; Li, liver. Scale bar: 50 µm.

    Journal: Nature

    Article Title: LUBAC is essential for embryogenesis by preventing cell death and enabling haematopoiesis

    doi: 10.1038/s41586-018-0064-8

    Figure Lengend Snippet: HOIL-1 deficiency causes embryonic lethality at mid-gestation due to TNFR1-mediated endothelial cell death a, Mendelian frequencies obtained from inter-crossing Hoil-1 +/- mice, *: dead embryos. b, Representative images of embryos from E9.5 to E11.5 quantified in (a), *: poor yolk sac vascularisation. Scale bar: 2 mm. c, Representative images of yolk sac vascularisation (PECAM-1, red) and cell death (cleaved (cl.) Caspase-3 staining, green) at E10.5 (top panel) ( n =4 yolk-sacs/genotype), and whole-mount TUNEL staining (bottom, panel) ( n =2 yolk-sacs/genotype). Scale bar: 50 µm. d, g, Quantification of branching points (g) and cleaved Caspase-3 positive cells (g). Mean ± s.e.m. values and P values from unpaired two-tailed t -tests are shown. e, Representative images of embryos at E10.5 ( n =14 Hoil-1 fl/wt Tie2-Cre+ and n =7 Hoil-1 fl/fl Tie2-Cre+ embryos, top panel). *: poor yolk sac vascularisation. Scale bar: 2 mm. Yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (middle panel). Scale bar: 50 µm. Yolk sac whole-mount TUNEL staining ( n =6 Hoil-1 fl/wt Tie2-Cre+ and n =2 Hoil-1 fl/fl Tie2-Cre+ yolk-sacs/genotype , bottom panel). f , Representative images of embryos at E15.5 (top panel, n =6 Tnfr1 -/- Hoil-1 -/- and n =19 Tnfr1 -/- Hoil-1 +/- embryos), scale bar: 2 mm, and yolk sac vascularisation (PECAM-1, red) and apoptosis (cleaved Caspase-3, green) (bottom panel), Scale bar: 50 µm. h, Representative images of H E staining on whole-embryo paraffin sections ( n =3 embryos/genotype). *: pericardial effusion, arrows; congested vessels. H, heart; L, lung; Li, liver. Scale bar: 50 µm.

    Article Snippet: Yolk sacs were stained with antibodies against PECAM-1 (BD Biosciences, 5533370 Clone MEC13.3) and cleaved caspase-3 (Cell Signaling, 9664), followed by staining with secondary antibodies, Alexa Fluor 594 Goat anti-Rat IgG and Alexa Fluor 488 Goat anti-Rabbit IgG (Invitrogen, A-11007 and A-11034, respectively), and analysed by fluorescent microscopy.

    Techniques: Mouse Assay, Staining, TUNEL Assay, Two Tailed Test

    AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.

    Journal: Scientific Reports

    Article Title: Silver nanoparticles have lethal and sublethal adverse effects on development and longevity by inducing ROS-mediated stress responses

    doi: 10.1038/s41598-018-20728-z

    Figure Lengend Snippet: AgNP-induced occurrence of systemic apoptosis. Immunofluorescent signals of the apoptotic biomarker “active caspase 3” in multiple tissues (brain, salivary gland, wing disc and gut) dissected from AgNP-exposed 3 rd -instar wandering larvae.

    Article Snippet: The rabbit anti-cleaved caspase 3 (1:250, Cell Signaling Technology, MA, USA) and rabbit anti-phosphorylated H2AX (1:500, Rockland, PA, USA) primary antibodies were used to quantitate the levels of AgNP-induced apoptosis and DNA damage (double-stranded breaks), respectively.

    Techniques: Biomarker Assay