anti monoubiquityl histone h2b lys 120  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti monoubiquityl histone h2b lys 120
    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in <t>H2B</t> monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Anti Monoubiquityl Histone H2b Lys 120, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes"

    Article Title: Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes

    Journal: Communications Biology

    doi: 10.1038/s42003-023-04424-x

    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Figure Legend Snippet: a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).

    Techniques Used: Mutagenesis, Two Tailed Test

    anti ubiquity histone h2b lys120  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti ubiquity histone h2b lys120
    Anti Ubiquity Histone H2b Lys120, 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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    anti histone h2b antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti histone h2b antibody
    Anti Histone H2b Antibody, 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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    anti monoubiquityl histone h2b lys 120  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti monoubiquityl histone h2b lys 120
    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in <t>H2B</t> monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Anti Monoubiquityl Histone H2b Lys 120, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes"

    Article Title: Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes

    Journal: Communications Biology

    doi: 10.1038/s42003-023-04424-x

    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Figure Legend Snippet: a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).

    Techniques Used: Mutagenesis, Two Tailed Test

    histone h2b  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc histone h2b
    Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone <t>H2B</t> antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.
    Histone H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A maladaptive feedback mechanism between the extracellular matrix and cytoskeleton contributes to hypertrophic cardiomyopathy pathophysiology"

    Article Title: A maladaptive feedback mechanism between the extracellular matrix and cytoskeleton contributes to hypertrophic cardiomyopathy pathophysiology

    Journal: Communications Biology

    doi: 10.1038/s42003-022-04278-9

    Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone H2B antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.
    Figure Legend Snippet: Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone H2B antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.

    Techniques Used: Western Blot, Expressing, Functional Assay

    histone h2b d2h6 rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc histone h2b d2h6 rabbit mab
    Histone H2b D2h6 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti histone h2b  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti histone h2b
    ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone <t>H2B</t> (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Anti Histone H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "ZBTB18 restricts chromatin accessibility and prevents transcriptional adaptations that drive metastasis"

    Article Title: ZBTB18 restricts chromatin accessibility and prevents transcriptional adaptations that drive metastasis

    Journal: Science Advances

    doi: 10.1126/sciadv.abq3951

    ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone H2B (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Figure Legend Snippet: ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone H2B (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Techniques Used: RNA Sequencing Assay, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

    h2b k120ub antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc h2b k120ub antibody
    SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant <t>H2B-Ub</t> nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM <t>H2B</t> ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.
    H2b K120ub Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The SAGA HAT module is tethered by its SWIRM domain and modulates activity of the SAGA DUB module"

    Article Title: The SAGA HAT module is tethered by its SWIRM domain and modulates activity of the SAGA DUB module

    Journal: bioRxiv

    doi: 10.1101/2022.12.29.522244

    SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant H2B-Ub nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM H2B ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.
    Figure Legend Snippet: SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant H2B-Ub nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM H2B ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.

    Techniques Used: Activity Assay, Quantitation Assay, Western Blot, Recombinant

    h2b  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc h2b
    H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti histone h2b  (Cell Signaling Technology Inc)


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  • 95

    Structured Review

    Cell Signaling Technology Inc rabbit anti histone h2b
    Antibodies, conjugates and immunoreagents.
    Rabbit Anti Histone H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Immunomodulatory Effects of Bone Marrow-Derived Mesenchymal Stem Cells on Pro-Inflammatory Cytokine-Stimulated Human Corneal Epithelial Cells"

    Article Title: Immunomodulatory Effects of Bone Marrow-Derived Mesenchymal Stem Cells on Pro-Inflammatory Cytokine-Stimulated Human Corneal Epithelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0101841

    Antibodies, conjugates and immunoreagents.
    Figure Legend Snippet: Antibodies, conjugates and immunoreagents.

    Techniques Used: Purification

    A to C shows FCM analysis of HCE-T cells treated with BMS-345541 (5 µM), a NF-κB pathway inhibitor, significantly inhibited IFN-γ (100 U/ml) and TNF (100 U/ml) induced ICAM-1 (A), HLA-ABC (B), and HLA-DR (C) expression. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05, **: p<0.01. D and E (upper panel) show representative bands of western blots for NF-κB p65 (65 kDa) in nuclear extracts (NE, D) and cytoplasmic extracts (CE, E). Histone H2B (15 kDa) and GAPDH (37 kDa) were used as the loading controls for nuclear and cytoplasmic extracts, respectively. Protein was prepared from HCE-T cells with or without IFN-γ/TNF stimulation, or HCE-T/MSC co-cultured with 24 h and 48 h IFN-γ/TNF treatment. The lower panel shows the fold changes of NF-κB within nuclear extracts and cytoplasmic extracts compared with the untreated group, using Histone H2B and GAPDH as loading controls. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05. F to I show representative immunocytochemistry images. Untreated HCE-T cells displayed weak NF-κB nuclear and cytoplasmic staining (F); NF-κB expression showed marked nuclear and perinuclear translocation after 24 h IFN-γ/TNF stimulation of HCE-T monocultures (G); NF-κB perinuclear and nuclear translocation decreased at 24 h in IFN-γ/TNF in stimulated HCECs/MSC co-cultures (H); HCE-T monolayer showed no obvious staining with rabbit IgG (I). The “+” or “−“ symbol denotes the presence or absence of the denoted treatment or cell type at the left.
    Figure Legend Snippet: A to C shows FCM analysis of HCE-T cells treated with BMS-345541 (5 µM), a NF-κB pathway inhibitor, significantly inhibited IFN-γ (100 U/ml) and TNF (100 U/ml) induced ICAM-1 (A), HLA-ABC (B), and HLA-DR (C) expression. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05, **: p<0.01. D and E (upper panel) show representative bands of western blots for NF-κB p65 (65 kDa) in nuclear extracts (NE, D) and cytoplasmic extracts (CE, E). Histone H2B (15 kDa) and GAPDH (37 kDa) were used as the loading controls for nuclear and cytoplasmic extracts, respectively. Protein was prepared from HCE-T cells with or without IFN-γ/TNF stimulation, or HCE-T/MSC co-cultured with 24 h and 48 h IFN-γ/TNF treatment. The lower panel shows the fold changes of NF-κB within nuclear extracts and cytoplasmic extracts compared with the untreated group, using Histone H2B and GAPDH as loading controls. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05. F to I show representative immunocytochemistry images. Untreated HCE-T cells displayed weak NF-κB nuclear and cytoplasmic staining (F); NF-κB expression showed marked nuclear and perinuclear translocation after 24 h IFN-γ/TNF stimulation of HCE-T monocultures (G); NF-κB perinuclear and nuclear translocation decreased at 24 h in IFN-γ/TNF in stimulated HCECs/MSC co-cultures (H); HCE-T monolayer showed no obvious staining with rabbit IgG (I). The “+” or “−“ symbol denotes the presence or absence of the denoted treatment or cell type at the left.

    Techniques Used: Expressing, Western Blot, Cell Culture, Immunocytochemistry, Staining, Translocation Assay

    ntcp  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc ntcp
    The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, <t>Ntcp)</t> and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 <t>and</t> <t>Bsep),</t> and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.
    Ntcp, 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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    Images

    1) Product Images from "Danning tablets attenuates α-naphthylisothiocyanate-induced cholestasis by modulating the expression of transporters and metabolic enzymes"

    Article Title: Danning tablets attenuates α-naphthylisothiocyanate-induced cholestasis by modulating the expression of transporters and metabolic enzymes

    Journal: BMC Complementary and Alternative Medicine

    doi: 10.1186/1472-6882-14-249

    The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, Ntcp) and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 and Bsep), and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.
    Figure Legend Snippet: The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, Ntcp) and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 and Bsep), and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Techniques Used: Quantitative RT-PCR, Isolation, Expressing

    The hepatic protein levels of transporters Oatp1, Ntcp, Mrp3, Mdr2 and Bsep were determined by Western blot (n = 3 for each group). Quantification of the protein expressions was performed by densitometric analysis of the blots following normalization to Na + /K + -ATPase expression. Each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.
    Figure Legend Snippet: The hepatic protein levels of transporters Oatp1, Ntcp, Mrp3, Mdr2 and Bsep were determined by Western blot (n = 3 for each group). Quantification of the protein expressions was performed by densitometric analysis of the blots following normalization to Na + /K + -ATPase expression. Each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Techniques Used: Western Blot, Expressing

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    Cell Signaling Technology Inc anti monoubiquityl histone h2b lys 120
    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in <t>H2B</t> monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Anti Monoubiquityl Histone H2b Lys 120, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti ubiquity histone h2b lys120
    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in <t>H2B</t> monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
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    Cell Signaling Technology Inc anti histone h2b antibody
    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in <t>H2B</t> monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).
    Anti Histone H2b Antibody, 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 histone h2b
    Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone <t>H2B</t> antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.
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    Cell Signaling Technology Inc histone h2b d2h6 rabbit mab
    Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone <t>H2B</t> antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.
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    Cell Signaling Technology Inc anti histone h2b
    ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone <t>H2B</t> (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Anti Histone H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc h2b k120ub antibody
    SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant <t>H2B-Ub</t> nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM <t>H2B</t> ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.
    H2b K120ub Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc h2b
    SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant <t>H2B-Ub</t> nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM <t>H2B</t> ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.
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    Cell Signaling Technology Inc rabbit anti histone h2b
    Antibodies, conjugates and immunoreagents.
    Rabbit Anti Histone H2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc ntcp
    The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, <t>Ntcp)</t> and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 <t>and</t> <t>Bsep),</t> and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.
    Ntcp, 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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    Image Search Results


    a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).

    Journal: Communications Biology

    Article Title: Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes

    doi: 10.1038/s42003-023-04424-x

    Figure Lengend Snippet: a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).

    Article Snippet: 5 µl of the crude chromatin extracts were saved for use as an input control, and 100 µl were added into the microwell immobilized with the antibodies (non-specific rabbit IgG Isotype Control (Invitrogen, WB317638, 1 µg) as the negative control, specific rabbit anti-His-tag antibody (GenScript, A00174-40, 2.5 µg) or anti-monoubiquityl-histone H2B (Lys-120) (5546S, Cell Signaling Technology, Inc., 2.5 µg).

    Techniques: Mutagenesis, Two Tailed Test

    Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone H2B antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.

    Journal: Communications Biology

    Article Title: A maladaptive feedback mechanism between the extracellular matrix and cytoskeleton contributes to hypertrophic cardiomyopathy pathophysiology

    doi: 10.1038/s42003-022-04278-9

    Figure Lengend Snippet: Immunoblot analysis of L-type calcium channel ( a , b ) and β 1 integrin ( c , d ) protein expression performed on total heart homogenate pooled from groups of 5 pre- (10–15-wk-old) or post-cardiomyopathic (30–50-week-old) cTnI-G203S mice and age-matched wt counterparts. Representative immunoblots probed with L-type calcium channel α 1C subunit (Ca V 1.2, a ) or β 1 integrin ( c ) antibody, then GAPDH monoclonal antibody. Densitometry analysis of Ca V 1.2 ( b ) or β 1 integrin ( d ) protein expression, normalized to associated GAPDH expression. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( b ) or Kruskal-Wallis test ( d ) determined statistical significance. Densitometry analysis of relative mTOR expression (calculated as Phospho-mTOR/Total mTOR) performed on cytoplasmic ( e ) and nuclear ( f ) fractions pooled from groups of five pre- or post-cardiomyopathic cTnI-G203S mice and age-matched wt counterparts. β-tubulin and histone H2B antibodies were used as loading controls for cytoplasmic and nuclear fractions respectively. n = number of technical repeats. A Browne-Forsythe and Welch ANOVA ( e ) or Kruskal-Wallis test ( f ) determined statistical significance. g Schematic indicating structural-functional link between the L-type calcium channel, cytoskeletal network, mitochondria, integrin and the extracellular matrix in wt and cTnI-G203S cardiac myocytes. A disrupted cytoskeletal architecture in cTnI-G203S cardiac myocytes may trigger a maladaptive feedback mechanism between increased cytoskeletal and extracellular matrix stiffness, resulting in a hypermetabolic mitochondrial state.

    Article Snippet: Histone H2B (D2H6, rabbit mAb, Cell Signaling Technology, 12364): Species: H, M, R, Mk; Applications: WB, IHC, ChiP; Validated by Cell Signaling Technology.

    Techniques: Western Blot, Expressing, Functional Assay

    ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone H2B (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Science Advances

    Article Title: ZBTB18 restricts chromatin accessibility and prevents transcriptional adaptations that drive metastasis

    doi: 10.1126/sciadv.abq3951

    Figure Lengend Snippet: ( A ) PCA of RNA-seq data indicates that E0771GFP, M11GFP, and M12GFP have distinct gene expression patterns ( n = 3 biological replicates per cell line). ( B ) Differential gene expression for M11GFP versus M12GFP and M11GFP versus E0771GFP. Only genes with significant differential expression ( P < 0.05) for both comparisons are shown. ( C ) Numbers of transcription factors with increased target gene expression ( P < 0.05) when comparing RNA-seq data obtained from E0771GFP, M11GFP, and M12GFP cell lines, as determined using GSEA analysis with the MSigDB C3 TFT (transcription factor target) collection. Genes with an FPKM of >1 were included in this analysis. ( D ) GSEA enrichment plot for ZBTB18 target gene expression in M11GFP cells compared with E0771GFP cells (left) and M12GFP cells (right). NES, normalized enrichment score. ( E ) Relative Zbtb18 mRNA levels in E0771GFP versus M11GFP (left; n = 3 biological replicates) and 67NR versus 4T1 (right; n = 4 biological replicates) cells, as determined by RT-qPCR. ( F ) ZBTB18 protein levels, detected by immunoblotting. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as loading control. Two biological replicates are shown. Right: Relative ZBTB18 expression levels calculated after normalization with GAPDH ( n = 4 biological replicates). ( G and H ) Relative expression levels of selected ZBTB18 target genes in E0771GFP and M11GFP (G) ( n = 3 biological replicates) or 67NR and 4T1 (H) ( n = 4 biological replicates) cells, as determined by RT-qPCR. ( I ) ZBTB18 protein expression levels, in whole-cell lysates and nuclear fractions, detected by immunoblotting. Nuclear histone H2B (H2B) and cytoplasmic GAPDH were used as controls. Relative ZBTB18 expression levels calculated after normalization with H2B are shown. ( J ) Immunofluorescence staining for ZBTB18 in E0771GFP and M11GFP cells. Cell nuclei are stained with DAPI (4′,6-diamidino-2-phenylindole). Scale bar, 50 μm. (E to H) Means ± SEM, unpaired two-sided t test. ns P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: The primary antibodies used were anti-ZBTB18 (ZNF238; Proteintech, #12714-1-AP; RRID:AB_2218388; Atlas Antibodies, HPA074019), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Millipore, #MAB374; RRID:AB_2107445), anti–histone H2B (H2B; Cell Signaling Technology, #12364;; RRID:AB_2714167), anti–phospho-Smad2 (Cell Signaling Technology, #3108, RRID:AB_490941), anti-Smad2/3 (Cell Signaling Technology, #8685; RRID:AB_10889933), and anti-TGFBR2 (Cell Signaling Technology, #79424; RRID:AB_2799933).

    Techniques: RNA Sequencing Assay, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

    SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant H2B-Ub nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM H2B ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.

    Journal: bioRxiv

    Article Title: The SAGA HAT module is tethered by its SWIRM domain and modulates activity of the SAGA DUB module

    doi: 10.1101/2022.12.29.522244

    Figure Lengend Snippet: SAGA’s HAT module regulates SAGA DUB activity independent of enzymatic activity. A: Quantitation of western blots of SAGA DUB activity on recombinant H2B-Ub nucleosomes (400 nM) under conditions of limiting SAGA enzyme (25 nM) with and without the SWIRM domain. B: Quantitation of western blots of SAGA DUB activity on HeLa nucleosomes with ~50 nM H2B ubiquitinated nucleosome. SAGA DUB activity is compared with and without the SWIRM domain, with excess SAGA, 100 nM. C: WT SAGA DUB (25 nM) activity on recombinant H2B-Ub nucleosomes (400 nM) in absence and presence of acetyl CoA (10 μ M). D: Quantitation of C and replicate.

    Article Snippet: Samples were visualized by western blot with an anti H2B-K120Ub antibody (Cell Signaling #5546).

    Techniques: Activity Assay, Quantitation Assay, Western Blot, Recombinant

    Antibodies, conjugates and immunoreagents.

    Journal: PLoS ONE

    Article Title: Immunomodulatory Effects of Bone Marrow-Derived Mesenchymal Stem Cells on Pro-Inflammatory Cytokine-Stimulated Human Corneal Epithelial Cells

    doi: 10.1371/journal.pone.0101841

    Figure Lengend Snippet: Antibodies, conjugates and immunoreagents.

    Article Snippet: Rabbit anti-histone H2B , 1∶1000 , WB , Cell Signaling, MA, USA.

    Techniques: Purification

    A to C shows FCM analysis of HCE-T cells treated with BMS-345541 (5 µM), a NF-κB pathway inhibitor, significantly inhibited IFN-γ (100 U/ml) and TNF (100 U/ml) induced ICAM-1 (A), HLA-ABC (B), and HLA-DR (C) expression. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05, **: p<0.01. D and E (upper panel) show representative bands of western blots for NF-κB p65 (65 kDa) in nuclear extracts (NE, D) and cytoplasmic extracts (CE, E). Histone H2B (15 kDa) and GAPDH (37 kDa) were used as the loading controls for nuclear and cytoplasmic extracts, respectively. Protein was prepared from HCE-T cells with or without IFN-γ/TNF stimulation, or HCE-T/MSC co-cultured with 24 h and 48 h IFN-γ/TNF treatment. The lower panel shows the fold changes of NF-κB within nuclear extracts and cytoplasmic extracts compared with the untreated group, using Histone H2B and GAPDH as loading controls. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05. F to I show representative immunocytochemistry images. Untreated HCE-T cells displayed weak NF-κB nuclear and cytoplasmic staining (F); NF-κB expression showed marked nuclear and perinuclear translocation after 24 h IFN-γ/TNF stimulation of HCE-T monocultures (G); NF-κB perinuclear and nuclear translocation decreased at 24 h in IFN-γ/TNF in stimulated HCECs/MSC co-cultures (H); HCE-T monolayer showed no obvious staining with rabbit IgG (I). The “+” or “−“ symbol denotes the presence or absence of the denoted treatment or cell type at the left.

    Journal: PLoS ONE

    Article Title: Immunomodulatory Effects of Bone Marrow-Derived Mesenchymal Stem Cells on Pro-Inflammatory Cytokine-Stimulated Human Corneal Epithelial Cells

    doi: 10.1371/journal.pone.0101841

    Figure Lengend Snippet: A to C shows FCM analysis of HCE-T cells treated with BMS-345541 (5 µM), a NF-κB pathway inhibitor, significantly inhibited IFN-γ (100 U/ml) and TNF (100 U/ml) induced ICAM-1 (A), HLA-ABC (B), and HLA-DR (C) expression. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05, **: p<0.01. D and E (upper panel) show representative bands of western blots for NF-κB p65 (65 kDa) in nuclear extracts (NE, D) and cytoplasmic extracts (CE, E). Histone H2B (15 kDa) and GAPDH (37 kDa) were used as the loading controls for nuclear and cytoplasmic extracts, respectively. Protein was prepared from HCE-T cells with or without IFN-γ/TNF stimulation, or HCE-T/MSC co-cultured with 24 h and 48 h IFN-γ/TNF treatment. The lower panel shows the fold changes of NF-κB within nuclear extracts and cytoplasmic extracts compared with the untreated group, using Histone H2B and GAPDH as loading controls. Data represents mean ± SEM of four separate experiments (n = 4). *: p<0.05. F to I show representative immunocytochemistry images. Untreated HCE-T cells displayed weak NF-κB nuclear and cytoplasmic staining (F); NF-κB expression showed marked nuclear and perinuclear translocation after 24 h IFN-γ/TNF stimulation of HCE-T monocultures (G); NF-κB perinuclear and nuclear translocation decreased at 24 h in IFN-γ/TNF in stimulated HCECs/MSC co-cultures (H); HCE-T monolayer showed no obvious staining with rabbit IgG (I). The “+” or “−“ symbol denotes the presence or absence of the denoted treatment or cell type at the left.

    Article Snippet: Rabbit anti-histone H2B , 1∶1000 , WB , Cell Signaling, MA, USA.

    Techniques: Expressing, Western Blot, Cell Culture, Immunocytochemistry, Staining, Translocation Assay

    The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, Ntcp) and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 and Bsep), and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Danning tablets attenuates α-naphthylisothiocyanate-induced cholestasis by modulating the expression of transporters and metabolic enzymes

    doi: 10.1186/1472-6882-14-249

    Figure Lengend Snippet: The effects of DNts on mRNA expressions of transporters and metabolic enzymes in the liver of ANIT-induced cholestasis rats. Relative hepatic mRNA levels of (A) basolateral uptake transporters (Oatp1, Ntcp) and basolateral efflux transporters (Mrp3), (B) basolateral efflux transporters (Mrp4, Ostα and Ostβ) (C) canalicular efflux transporters (Mrp2, Mdr2 and Bsep), and (D) bile acid-metabolizing enzymes (Cyp2b1, Ugt1a1 and Sult2a1) were determined by qRT-PCR in liver samples isolated from rats treated with vehicle, DNts, ANIT or DNts + ANIT (n = 3 for each group). Expression was normalized to Gapdh, and each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Article Snippet: Membranes were blocked in 5% skim milk and incubated with antibodies against Bsep (sc-25571, 1:200, Santa Cruz), Ntcp (#6522-1, 1:1000, epitomics), Oatp1 (LS-C113034-50, 1:1000, LifeSpan BioSciences), Mrp3(ab3375, 1:50, abcam), Mdr2 (SAB2100008, 1:1000, sigma) and Na + /K + -ATPase (#3010, 1:1000, Cell Signaling), respectively.

    Techniques: Quantitative RT-PCR, Isolation, Expressing

    The hepatic protein levels of transporters Oatp1, Ntcp, Mrp3, Mdr2 and Bsep were determined by Western blot (n = 3 for each group). Quantification of the protein expressions was performed by densitometric analysis of the blots following normalization to Na + /K + -ATPase expression. Each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Danning tablets attenuates α-naphthylisothiocyanate-induced cholestasis by modulating the expression of transporters and metabolic enzymes

    doi: 10.1186/1472-6882-14-249

    Figure Lengend Snippet: The hepatic protein levels of transporters Oatp1, Ntcp, Mrp3, Mdr2 and Bsep were determined by Western blot (n = 3 for each group). Quantification of the protein expressions was performed by densitometric analysis of the blots following normalization to Na + /K + -ATPase expression. Each bar represents the mean ± SD of three independent experiments. Significant difference between two groups. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. the ANIT-treated group.

    Article Snippet: Membranes were blocked in 5% skim milk and incubated with antibodies against Bsep (sc-25571, 1:200, Santa Cruz), Ntcp (#6522-1, 1:1000, epitomics), Oatp1 (LS-C113034-50, 1:1000, LifeSpan BioSciences), Mrp3(ab3375, 1:50, abcam), Mdr2 (SAB2100008, 1:1000, sigma) and Na + /K + -ATPase (#3010, 1:1000, Cell Signaling), respectively.

    Techniques: Western Blot, Expressing