rabbit anti phospho histone3 phis  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho histone3 phis
    Rabbit Anti Phospho Histone3 Phis, 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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    rabbit antibodies anti phospho histone3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit antibodies anti phospho histone3
    Rabbit Antibodies Anti Phospho Histone3, 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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    phospho histone3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3
    Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle <t>(Phospho-Histone3</t> immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).
    Phospho Histone3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Adaptation of NS cells growth and differentiation to high-throughput screening-compatible plates"

    Article Title: Adaptation of NS cells growth and differentiation to high-throughput screening-compatible plates

    Journal: BMC Neuroscience

    doi: 10.1186/1471-2202-11-7

    Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle (Phospho-Histone3 immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).
    Figure Legend Snippet: Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle (Phospho-Histone3 immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).

    Techniques Used: Cell Culture, Expressing

    phospho histone3 ph3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3 ph3
    A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 <t>(pH3)</t> immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.
    Phospho Histone3 Ph3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Spry1 and Spry4 Differentially Regulate Human Aortic Smooth Muscle Cell Phenotype via Akt/FoxO/Myocardin Signaling"

    Article Title: Spry1 and Spry4 Differentially Regulate Human Aortic Smooth Muscle Cell Phenotype via Akt/FoxO/Myocardin Signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0058746

    A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 (pH3) immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.
    Figure Legend Snippet: A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 (pH3) immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.

    Techniques Used: Immunofluorescence, Staining, Immunohistochemistry, Expressing, Quantitative RT-PCR, Sequencing, Western Blot, Marker

    phospho histone3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3
    Phospho Histone3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho histone3/product/Cell Signaling Technology Inc
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    phospho histone3 immunostaining  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3 immunostaining
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    phospho histone3 ser10 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3 ser10 antibody
    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for <t>phospho-histone3</t> (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.
    Phospho Histone3 Ser10 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho histone3 ser10 antibody/product/Cell Signaling Technology Inc
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    1) Product Images from "Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion"

    Article Title: Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion

    Journal: Nature Genetics

    doi: 10.1038/s41588-022-01191-z

    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for phospho-histone3 (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.
    Figure Legend Snippet: a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for phospho-histone3 (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.

    Techniques Used: Mutagenesis, Expressing, CRISPR, Plasmid Preparation, Two Tailed Test, Whisker Assay, Injection, Knock-Out, Staining, Immunohistochemistry, Transplantation Assay, Cell Counting

    rabbit antibodies anti phospho histone3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit antibodies anti phospho histone3
    Cell proliferation and viability of genetic combinations affecting wing size. (A-B) Wing phenotype (A and B), expression of <t>phospho-Histone3</t> (pH3; red in A’ and B’) and cleaved-Dcp1 (DcpI*, white in A’’ and B’’) in control UAS-Dicer2; nub-Gal4/UAS-GFP third instar wing discs grown at 25°C (A–A’’) and 29°C (B–B’’). (C–F) Wing phenotype (C–F), expression of phospho-Histone3 (pH3; red in C’–F’), and cleaved-Dcp1 (DcpI*, white in C’’–F’’) in the genetic combinations UAS-Dicer2; nub-Gal4/UAS-fab1-RNAi (C–C’’), UAS-Dicer2; nub-Gal4/UAS-cdc7-RNAi (D–D’’), UAS-Dicer2; nub-Gal4/UAS-CG14297-RNAi (E–E’’), and UAS-Dicer2; nub-Gal4/UAS-Takl2-RNAi (F–F’’). Below each wing is indicated the percentage of wing size (Size), cell size (cell size), and wing cell number (cell no.) modification for each genetic combination compared to their control UAS-Dicer2; nub-Gal4/UAS-GFP wings.
    Rabbit Antibodies Anti Phospho Histone3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Functional requirements of protein kinases and phosphatases in the development of the Drosophila melanogaster wing"

    Article Title: Functional requirements of protein kinases and phosphatases in the development of the Drosophila melanogaster wing

    Journal: G3: Genes|Genomes|Genetics

    doi: 10.1093/g3journal/jkab348

    Cell proliferation and viability of genetic combinations affecting wing size. (A-B) Wing phenotype (A and B), expression of phospho-Histone3 (pH3; red in A’ and B’) and cleaved-Dcp1 (DcpI*, white in A’’ and B’’) in control UAS-Dicer2; nub-Gal4/UAS-GFP third instar wing discs grown at 25°C (A–A’’) and 29°C (B–B’’). (C–F) Wing phenotype (C–F), expression of phospho-Histone3 (pH3; red in C’–F’), and cleaved-Dcp1 (DcpI*, white in C’’–F’’) in the genetic combinations UAS-Dicer2; nub-Gal4/UAS-fab1-RNAi (C–C’’), UAS-Dicer2; nub-Gal4/UAS-cdc7-RNAi (D–D’’), UAS-Dicer2; nub-Gal4/UAS-CG14297-RNAi (E–E’’), and UAS-Dicer2; nub-Gal4/UAS-Takl2-RNAi (F–F’’). Below each wing is indicated the percentage of wing size (Size), cell size (cell size), and wing cell number (cell no.) modification for each genetic combination compared to their control UAS-Dicer2; nub-Gal4/UAS-GFP wings.
    Figure Legend Snippet: Cell proliferation and viability of genetic combinations affecting wing size. (A-B) Wing phenotype (A and B), expression of phospho-Histone3 (pH3; red in A’ and B’) and cleaved-Dcp1 (DcpI*, white in A’’ and B’’) in control UAS-Dicer2; nub-Gal4/UAS-GFP third instar wing discs grown at 25°C (A–A’’) and 29°C (B–B’’). (C–F) Wing phenotype (C–F), expression of phospho-Histone3 (pH3; red in C’–F’), and cleaved-Dcp1 (DcpI*, white in C’’–F’’) in the genetic combinations UAS-Dicer2; nub-Gal4/UAS-fab1-RNAi (C–C’’), UAS-Dicer2; nub-Gal4/UAS-cdc7-RNAi (D–D’’), UAS-Dicer2; nub-Gal4/UAS-CG14297-RNAi (E–E’’), and UAS-Dicer2; nub-Gal4/UAS-Takl2-RNAi (F–F’’). Below each wing is indicated the percentage of wing size (Size), cell size (cell size), and wing cell number (cell no.) modification for each genetic combination compared to their control UAS-Dicer2; nub-Gal4/UAS-GFP wings.

    Techniques Used: Expressing, Modification

    anti phospho histone3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho histone3
    Anti Phospho Histone3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho histone3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone3 ser10
    Phospho Histone3 Ser10, 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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    Cell Signaling Technology Inc rabbit anti phospho histone3 phis
    Rabbit Anti Phospho Histone3 Phis, 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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    Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle <t>(Phospho-Histone3</t> immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).
    Phospho Histone3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho histone3 ph3
    A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 <t>(pH3)</t> immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.
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    A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 <t>(pH3)</t> immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.
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    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for <t>phospho-histone3</t> (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.
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    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for <t>phospho-histone3</t> (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.
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    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for <t>phospho-histone3</t> (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.
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    Image Search Results


    Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle (Phospho-Histone3 immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).

    Journal: BMC Neuroscience

    Article Title: Adaptation of NS cells growth and differentiation to high-throughput screening-compatible plates

    doi: 10.1186/1471-2202-11-7

    Figure Lengend Snippet: Adaptation of culture conditions of aNS-1 cells to microplates . aNS-1 cells can be cultured in 96- ( A ) and 384-well ( B ) plates, maintaining the expression of proper NS cells markers (Nestin, Vimentin, BLBP, Olig2), without exiting the cell cycle (Phospho-Histone3 immunoreactivity, P-H3), without differentiation (beta III-tubulin, MAP2 and GFAP absence) or cell death (cleaved caspase-3 absence).

    Article Snippet: Primary antibodies used in this work are: anti beta III-tubulin (1:1000; Promega); anti BLBP (1:1000; Chemicon); anti cleaved caspase-3 (1:100; Cell Signaling); anti GFAP (1:750; DAKO); anti MAP2 (1:500; Chemicon); anti Nestin (1:250; Chemicon); anti Olig2 (1:500; Chemicon); anti Phospho Histone3 (1:100; Cell Signaling); anti Vimentin (1:100; Developmental Studies Hybridoma Bank).

    Techniques: Cell Culture, Expressing

    A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 (pH3) immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.

    Journal: PLoS ONE

    Article Title: Spry1 and Spry4 Differentially Regulate Human Aortic Smooth Muscle Cell Phenotype via Akt/FoxO/Myocardin Signaling

    doi: 10.1371/journal.pone.0058746

    Figure Lengend Snippet: A) Co-immunofluorescence staining of Spry1, Spry4 and ACTA2 on mouse artery sections show that both Spry1 and Spry4 are expressed in normal arterial smooth muscle cells. Images are from original 400× magnification. B) Immunohistochemistry of Spry1 and Spry4 on human atherosclerotic artery samples showed the expression of Spry1 and Spry4 in artery media smooth muscle cells, and a decreased expression of Spry1 and Spry4 in neointima cells. Images are from original 200× magnification. C) Quantification of RT-qPCR from three independent experiments shows efficacy of knockdown (kd) of Spry1 and Spry4 mRNA by their respective shRNAs compared to a non-targeting sequence (NT). D) Quantification of RT-qPCR from three independent experiments shows that knockdown of Spry1 decreases, whereas knockdown of Spry4 increase VSMC differentiation gene expression at the mRNA level. E) Representative immunoblot showing that knockdown of Spry1 results in a decrease in total Akt and ERK proteins levels, as well as pAkt and pERK. The expression of VSMC marker genes SMTN-B and SM22α is decreased by knockdown of Spry1, whereas knockdown of Spry4 increases pAkt and pERK levels as well as SMTN-B and SM22α. F) Quantification of VSMC marker protein levels from at least three independent immunoblotting experiments. G) Immunofluorescence staining shows down-regulation of SM22α following knockdown of Spry1. Images are from original 400× magnification. H) Quantification of phospho-histone 3 (pH3) immunofluorescence staining from triplicate experiments shows that knockdown of Spry1 reduces pH3 staining, while knockdown of Spry4 increases pH3 staining. Note: a: adventia, m: media, i: intima l: lumen; **: p<0.01, *: p<0.05.

    Article Snippet: For phospho-histone3 (pH3) and FoxO3a immunostaining, cells were fixed in 4% PFA for 10 min, stained with anti-pH3 (Upstate, 1∶200) or anti-FoxO3a (Cell Signaling, 1∶50) followed by FITC-anti-rabbit antibody (BioRad).

    Techniques: Immunofluorescence, Staining, Immunohistochemistry, Expressing, Quantitative RT-PCR, Sequencing, Western Blot, Marker

    a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for phospho-histone3 (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.

    Journal: Nature Genetics

    Article Title: Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion

    doi: 10.1038/s41588-022-01191-z

    Figure Lengend Snippet: a , Schematic representation of the generation of genetically engineered primary melanomas in adult zebrafish (Methods). b , Proportion of nectin1a and nectin1b mutant alleles in 65 primary zebrafish melanomas expressing a CRISPR vector targeting nectin1a and nectin1b . Bar represents median; paired two-tailed t-test. c , Box-and-whisker plot representing the number of nectin1a and nectin1b mutant alleles per primary zebrafish melanoma described in panel b (min-max, 1st and 4th quartiles, median, n = 65; paired two-tailed t-test). d , Type and proportion of nectin1a and nectin1b mutant alleles in 10 representative primary zebrafish melanomas. D, deletion, I, insertion; numbers denote affected base pairs. The last section in each bar represents all other minor alleles. e , Tumor-free survival curves of Tg(mitf:BRAF V600E );tp53 −/− zebrafish injected with vectors targeting either nectin1 or a control gene (pooled data of three independent experiments; log-rank test). f , Representative images of sections of control - or nectin1 -knockout (KO) primary zebrafish melanomas, stained for phospho-histone3 (p-H3) by immunohistochemistry (IHC). Scale bar, 20 μm. g , Quantification of overall p-H3 staining intensity in 8 nectin1 -wildype and 10 nectin1 -knockout primary zebrafish melanomas (mean ± s.d.; two-tailed t -test). NS, not significant. h , Pictures of organs presenting with disseminated tumor cells (arrowheads) in hematoxylin/eosin-stained sections of 16-week-old zebrafish bearing nectin1 -knockout melanoma. Scale bar, 20 μm. i , Number of 16-week-old zebrafish with disseminated melanoma cells in the indicated organs. j , Quantification of organ involvement in 16-week-old zebrafish bearing control ( n = 11) or nectin1 -knockout ( n = 10) melanoma (mean ± s.d.). Two-tailed t-test. k , Schematic representation of the melanoma spreading assay in adult zebrafish (top) and representative images of casper zebrafish 7 or 21 days after transplantation with control - or nectin1 -knockout melanoma cells (bottom). Insets show ×2 magnification views. Arrows indicate patches of disseminated melanoma cells. Scale bar, 1 cm. l , Quantification of the proportion of secondary recipients of control - ( n = 10) or nectin1 -knockout ( n = 9) tumors showing tumor spreading (mean ± s.d.; two-tailed t -test). m , Cell count of control - and nectin1 -knockout primary zebrafish melanoma lines in the presence or absence of zebrafish (zf) culture media. Data are presented as mean ± s.d. of three independent experiments.

    Article Snippet: Phospho-Histone3 (Ser10) antibody (#9701, Cell Signaling Technology) or phospho-FAK (Tyr397) antibody (44–624 G, Invitrogen) were run at 1:200 dilution using the Leica Biosystems Refine Detection Kit with citrate antigen retrieval, after bleaching of the melanin pigment.

    Techniques: Mutagenesis, Expressing, CRISPR, Plasmid Preparation, Two Tailed Test, Whisker Assay, Injection, Knock-Out, Staining, Immunohistochemistry, Transplantation Assay, Cell Counting