rabbit anti et b  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti et b
    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections <t>with</t> <t>antibodies</t> against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for <t>ET</t> <t>B</t> ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.
    Rabbit Anti Et B, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti et b/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti et b - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia"

    Article Title: Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0041852

    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.
    Figure Legend Snippet: Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.

    Techniques Used: Immunohistochemical staining, Staining

    Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.
    Figure Legend Snippet: Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.

    Techniques Used: Western Blot, Expressing

    Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.
    Figure Legend Snippet: Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.

    Techniques Used: Concentration Assay, Expressing, Immunohistochemical staining, Staining

    rabbit anti et b  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
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  • 94

    Structured Review

    Alomone Labs rabbit anti et b
    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections <t>with</t> <t>antibodies</t> against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for <t>ET</t> <t>B</t> ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.
    Rabbit Anti Et B, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti et b/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti et b - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia"

    Article Title: Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0041852

    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.
    Figure Legend Snippet: Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.

    Techniques Used: Immunohistochemical staining, Staining

    Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.
    Figure Legend Snippet: Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.

    Techniques Used: Western Blot, Expressing

    Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.
    Figure Legend Snippet: Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.

    Techniques Used: Concentration Assay, Expressing, Immunohistochemical staining, Staining

    rabbit anti ednrb antibody  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti ednrb antibody
    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
    Rabbit Anti Ednrb Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti ednrb antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ednrb antibody - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage"

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    Journal: Scientific Reports

    doi: 10.1038/s41598-022-20497-w

    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
    Figure Legend Snippet: Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Techniques Used: Injection

    mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.
    Figure Legend Snippet: mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Techniques Used: Expressing, Injection, Quantitative RT-PCR, Isolation, Cell Culture

    Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.
    Figure Legend Snippet: Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Techniques Used: Expressing, Labeling, Injection, Western Blot, Staining, Software

    EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.
    Figure Legend Snippet: EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Techniques Used: Expressing, Cell Culture, Labeling, Western Blot

    rabbit anti ednrb  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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  • 94

    Structured Review

    Alomone Labs rabbit anti ednrb
    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
    Rabbit Anti Ednrb, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti ednrb/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ednrb - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage"

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    Journal: Scientific Reports

    doi: 10.1038/s41598-022-20497-w

    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
    Figure Legend Snippet: Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Techniques Used: Injection

    mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.
    Figure Legend Snippet: mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Techniques Used: Expressing, Injection, Quantitative RT-PCR, Isolation, Cell Culture

    Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.
    Figure Legend Snippet: Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Techniques Used: Expressing, Labeling, Injection, Western Blot, Staining, Software

    EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.
    Figure Legend Snippet: EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Techniques Used: Expressing, Cell Culture, Labeling, Western Blot

    polyclonal rabbit anti endothelin et b receptor antibody  (Alomone Labs)


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    Alomone Labs polyclonal rabbit anti endothelin et b receptor antibody
    Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of <t>endothelin-1</t> (ET-1) after <t>ET</t> <t>B</t> receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.
    Polyclonal Rabbit Anti Endothelin Et B Receptor Antibody, supplied by Alomone Labs, 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 "Myocardial ischemia-reperfusion enhances transcriptional expression of endothelin-1 and vasoconstrictor ET B receptors via the protein kinase MEK-ERK1/2 signaling pathway in rat"

    Article Title: Myocardial ischemia-reperfusion enhances transcriptional expression of endothelin-1 and vasoconstrictor ET B receptors via the protein kinase MEK-ERK1/2 signaling pathway in rat

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174119

    Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of endothelin-1 (ET-1) after ET B receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.
    Figure Legend Snippet: Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of endothelin-1 (ET-1) after ET B receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.

    Techniques Used: Concentration Assay

    ednrb rabbit  (Alomone Labs)


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    Alomone Labs ednrb rabbit
    Antibodies.
    Ednrb Rabbit, supplied by Alomone Labs, 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 "A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions"

    Article Title: A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions

    Journal: F1000Research

    doi: 10.12688/f1000research.7142.2

    Antibodies.
    Figure Legend Snippet: Antibodies.

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    rabbit anti et b r  (Alomone Labs)


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    Alomone Labs rabbit anti et b r
    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the <t>ET</t> <t>B</t> <t>R</t> mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative <t>immunofluorescence</t> <t>staining</t> on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).
    Rabbit Anti Et B R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti et b r/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    1) Product Images from "Human Cytomegalovirus Up-Regulates Endothelin Receptor Type B: Implication for Vasculopathies?"

    Article Title: Human Cytomegalovirus Up-Regulates Endothelin Receptor Type B: Implication for Vasculopathies?

    Journal: Open Forum Infectious Diseases

    doi: 10.1093/ofid/ofv155

    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the ET B R mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative immunofluorescence staining on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).
    Figure Legend Snippet: Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the ET B R mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative immunofluorescence staining on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).

    Techniques Used: Expressing, Infection, Polymerase Chain Reaction, Irradiation, Standard Deviation, Immunofluorescence, Staining, Fluorescence, Software

    Human cytomegalovirus (HCMV) infection alters endothelin receptor type B (ET B R) cellular distribution, induces surface expression of ET B R and its effect on endothelin receptor type A (ET A R). (A) Representative immunofluorescence staining on HCMV-infected endothelial cells at 3 days postinfection to illustrate the subcellular distribution of ET B R in uninfected and infected cells. Arrow shows increased ET B R protein expression in the perinuclear region. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). (B) Representative immunofluorescence staining of ET B R and giantin, a Golgi marker (top panel), or PDI, an endoplasmic reticulum marker (bottom panel). Nucleus was stained with DAPI (blue). Flow cytometry analysis on the effects of HCMV-infected endothelial cells (human umbilical vein cells [HUVECs]) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs] and human aortic SMCs [hSMCs]) on ET B R at indicated days postinfection (dpi). (C) The effects of different multiplicity of infection (moi) of HCMV on ET B R surface expression at 1, 3, and 5 dpi (left panel) in HUVECs. The HCMV IE is shown on the right panel. Y-axis denotes log intensity of ET B R (left panel) or IE (right panel), whereas x-axis is side scatter in linear scale (SS Lin). (D) The effects of ultraviolet (UV)-irradiated HCMV on ET B R surface expression in HUVECs. Little or no IE was detected with the UV treatment (bottom row). (E) The effects of UV-irradiated HCMV (middle row) or HCMV (bottom row) on ET B R surface expression in SMCs, hPASMCs (left column) and hSMCs (right column) at 5 dpi. Y-axis denotes log intensity of ET B R (left panel), whereas x-axis is SS Lin. The effects of HCMV on ET A R mRNA level in smooth muscle cells, hPASMC (F) and hSMC (G) (MOI; biological replicates, n = 3 with technical duplicates for each polymerase chain reaction; values are mean ± standard deviation; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001).
    Figure Legend Snippet: Human cytomegalovirus (HCMV) infection alters endothelin receptor type B (ET B R) cellular distribution, induces surface expression of ET B R and its effect on endothelin receptor type A (ET A R). (A) Representative immunofluorescence staining on HCMV-infected endothelial cells at 3 days postinfection to illustrate the subcellular distribution of ET B R in uninfected and infected cells. Arrow shows increased ET B R protein expression in the perinuclear region. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). (B) Representative immunofluorescence staining of ET B R and giantin, a Golgi marker (top panel), or PDI, an endoplasmic reticulum marker (bottom panel). Nucleus was stained with DAPI (blue). Flow cytometry analysis on the effects of HCMV-infected endothelial cells (human umbilical vein cells [HUVECs]) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs] and human aortic SMCs [hSMCs]) on ET B R at indicated days postinfection (dpi). (C) The effects of different multiplicity of infection (moi) of HCMV on ET B R surface expression at 1, 3, and 5 dpi (left panel) in HUVECs. The HCMV IE is shown on the right panel. Y-axis denotes log intensity of ET B R (left panel) or IE (right panel), whereas x-axis is side scatter in linear scale (SS Lin). (D) The effects of ultraviolet (UV)-irradiated HCMV on ET B R surface expression in HUVECs. Little or no IE was detected with the UV treatment (bottom row). (E) The effects of UV-irradiated HCMV (middle row) or HCMV (bottom row) on ET B R surface expression in SMCs, hPASMCs (left column) and hSMCs (right column) at 5 dpi. Y-axis denotes log intensity of ET B R (left panel), whereas x-axis is SS Lin. The effects of HCMV on ET A R mRNA level in smooth muscle cells, hPASMC (F) and hSMC (G) (MOI; biological replicates, n = 3 with technical duplicates for each polymerase chain reaction; values are mean ± standard deviation; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001).

    Techniques Used: Infection, Expressing, Immunofluorescence, Staining, Marker, Flow Cytometry, Irradiation, Polymerase Chain Reaction, Standard Deviation

    Human cytomegalovirus (HCMV) antigens are closely associated with endothelin receptor type B (ET B R) in human carotid atherosclerotic plaques. Representative photomicrographs of immunohistochemistry and immunofluorescence (IF) staining on frozen human carotid atherosclerotic plaques. (A) Immunohistochemistry staining to show colocalization of HCMV immediate-early (IE) and ET B R in Von Willebrand factor (vWF)-positive cells as revealed by chromogen diaminobenzidine, brown products in a boxed area with consecutive sections (left column; right column is a higher magnification of the boxed areas). (B) The IF staining on a human carotid atherosclerotic plaque to illustrate the colocalization of HCMV-IE with ET B R in fibroblast-like cells. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of HCMV-IE or ET B R was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively. Twenty human carotid atherosclerotic plaques were examined.
    Figure Legend Snippet: Human cytomegalovirus (HCMV) antigens are closely associated with endothelin receptor type B (ET B R) in human carotid atherosclerotic plaques. Representative photomicrographs of immunohistochemistry and immunofluorescence (IF) staining on frozen human carotid atherosclerotic plaques. (A) Immunohistochemistry staining to show colocalization of HCMV immediate-early (IE) and ET B R in Von Willebrand factor (vWF)-positive cells as revealed by chromogen diaminobenzidine, brown products in a boxed area with consecutive sections (left column; right column is a higher magnification of the boxed areas). (B) The IF staining on a human carotid atherosclerotic plaque to illustrate the colocalization of HCMV-IE with ET B R in fibroblast-like cells. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of HCMV-IE or ET B R was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively. Twenty human carotid atherosclerotic plaques were examined.

    Techniques Used: Immunohistochemistry, Immunofluorescence, Staining

    antibody rabbit anti et b receptor  (Alomone Labs)


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    Alomone Labs antibody rabbit anti et b receptor
    Antibody Rabbit Anti Et B Receptor, supplied by Alomone Labs, 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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    antibody rabbit anti et b receptor  (Alomone Labs)


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    Alomone Labs antibody rabbit anti et b receptor
    Antibody Rabbit Anti Et B Receptor, supplied by Alomone Labs, 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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    rabbit anti et b receptor antibody  (Alomone Labs)


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    Alomone Labs rabbit anti et b receptor antibody
    Rabbit Anti Et B Receptor Antibody, supplied by Alomone Labs, 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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    rabbit anti et b receptor antibody  (Alomone Labs)


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    Alomone Labs rabbit anti et b receptor antibody
    Rabbit Anti Et B Receptor Antibody, supplied by Alomone Labs, 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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    Alomone Labs rabbit anti et b
    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections <t>with</t> <t>antibodies</t> against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for <t>ET</t> <t>B</t> ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.
    Rabbit Anti Et B, supplied by Alomone Labs, 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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    Alomone Labs rabbit anti ednrb antibody
    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
    Rabbit Anti Ednrb Antibody, supplied by Alomone Labs, 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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    Alomone Labs rabbit anti ednrb
    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.
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    Alomone Labs polyclonal rabbit anti endothelin et b receptor antibody
    Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of <t>endothelin-1</t> (ET-1) after <t>ET</t> <t>B</t> receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.
    Polyclonal Rabbit Anti Endothelin Et B Receptor Antibody, supplied by Alomone Labs, 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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    Alomone Labs ednrb rabbit
    Antibodies.
    Ednrb Rabbit, supplied by Alomone Labs, 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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    94
    Alomone Labs rabbit anti et b r
    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the <t>ET</t> <t>B</t> <t>R</t> mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative <t>immunofluorescence</t> <t>staining</t> on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).
    Rabbit Anti Et B R, supplied by Alomone Labs, 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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    94
    Alomone Labs antibody rabbit anti et b receptor
    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the <t>ET</t> <t>B</t> <t>R</t> mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative <t>immunofluorescence</t> <t>staining</t> on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).
    Antibody Rabbit Anti Et B Receptor, supplied by Alomone Labs, 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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    94
    Alomone Labs rabbit anti et b receptor antibody
    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the <t>ET</t> <t>B</t> <t>R</t> mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative <t>immunofluorescence</t> <t>staining</t> on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).
    Rabbit Anti Et B Receptor Antibody, supplied by Alomone Labs, 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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    Image Search Results


    Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.

    Journal: PLoS ONE

    Article Title: Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia

    doi: 10.1371/journal.pone.0041852

    Figure Lengend Snippet: Photomicrographs showing immunohistochemical stainings of anterior cerebral artery (ACA) sections with antibodies against endothelin type B (ET B ) ( A and B ) or endothelin typ A (ET A ) ( D and E ) receptors. Bar graphs show quantifications of staining intensities for ET B ( C ) and ET A ( F ) receptors. Two sections from each of 6 to 11 animals were analyzed. Data are presented as mean SEM in percentages of the mean staining intensity in control-operated (sham) animals. Significant differences between sham-operated and 15 minutes ischemia rats were determined using student´s t-test. *p< 0.05.

    Article Snippet: The membranes were then blocked for unspecific binding for 1 hour at room temperature in TBS-T buffer (TBS +1% Tween20 (sigma) containing 5% BSA and thereafter incubated over night at 4°C with primary antibodies: Rabbit anti-ET B (Alomone labs) diluted 1∶250, goat anti-5-HT 1B (Mybiosource) diluted 1∶250 or mouse anti-actin (Abcam) diluted 1∶15000.

    Techniques: Immunohistochemical staining, Staining

    Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.

    Journal: PLoS ONE

    Article Title: Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia

    doi: 10.1371/journal.pone.0041852

    Figure Lengend Snippet: Representative western blots and band intensity quantifications showing endothelin type B (ET B ) protein expression in basilar artery (BA) ( A ) and in pooled middle cerebral artery (MCA) and anterior cerebral artery (ACA) tissue ( B ), and 5-hydroxotryptamin type 1B (5-HT 1B ) protein expression in BA ( C ) and in pooled MCA and ACA tissue ( D ) from control-operated (sham) rats and rats subjected to 15 minutes of transient forebrain ischemia. The band recognised by the ET B receptor antibody was approximately 48 kDa and the band recognised by the 5-HT1B receptor antibody was approximately 41 kDa. Actin was used as a loading control. Data are expressed as mean ± SEM percentages of the mean band intensity in sham-operated animals and student’s t-test was used for statistical comparison. n = 5–13. *p<0.05.

    Article Snippet: The membranes were then blocked for unspecific binding for 1 hour at room temperature in TBS-T buffer (TBS +1% Tween20 (sigma) containing 5% BSA and thereafter incubated over night at 4°C with primary antibodies: Rabbit anti-ET B (Alomone labs) diluted 1∶250, goat anti-5-HT 1B (Mybiosource) diluted 1∶250 or mouse anti-actin (Abcam) diluted 1∶15000.

    Techniques: Western Blot, Expressing

    Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.

    Journal: PLoS ONE

    Article Title: Expressional Changes in Cerebrovascular Receptors after Experimental Transient Forebrain Ischemia

    doi: 10.1371/journal.pone.0041852

    Figure Lengend Snippet: Data for endothelin type B (ET B ) and 5-hydroxotryptamin type 1B (5-HT 1B ) receptor-mediated contractile function represent E max values for the first phase (E max-1 ) of the biphasic concentration-contraction curves for anterior cerebral arteries (ACA) stimulated with endothelin-1 (ET-1) and 5-carboxamidotryptamine (5-CT), respectively. Data for ET B and 5-HT 1B protein expression levels represent receptor subtype-specific immunohistochemical staining intensities in ACAs. Neurology score data represents pooled data from grip strength and rotating pole. All data are values for ischemia-induced rats (2, 10 or 15 minutes) terminated 48 hours after ischemia in percentages of corresponding values for control-operated (sham) rats. All data are expressed as mean ± SEM.

    Article Snippet: The membranes were then blocked for unspecific binding for 1 hour at room temperature in TBS-T buffer (TBS +1% Tween20 (sigma) containing 5% BSA and thereafter incubated over night at 4°C with primary antibodies: Rabbit anti-ET B (Alomone labs) diluted 1∶250, goat anti-5-HT 1B (Mybiosource) diluted 1∶250 or mouse anti-actin (Abcam) diluted 1∶15000.

    Techniques: Concentration Assay, Expressing, Immunohistochemical staining, Staining

    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

    Techniques: Injection

    mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

    Techniques: Expressing, Injection, Quantitative RT-PCR, Isolation, Cell Culture

    Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

    Techniques: Expressing, Labeling, Injection, Western Blot, Staining, Software

    EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

    Techniques: Expressing, Cell Culture, Labeling, Western Blot

    Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: Genes related to the excretion GO term and upregulated by NMDA injection and downregulated by KUS121 and KUS187 treatment.

    Article Snippet: The primary antibodies used to probe the blots were mouse anti-EDN1 (1:400; Abcam, Cambridge, UK), rabbit anti-EDNRB (1:400; Alomone Labs, Jerusalem, Israel), rabbit anti-AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-AKT (1:2000, Cell Signaling Technology), rabbit anti-ERK (1:20,000, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-phospho-ERK (1:1000, Cell Signaling Technology) and anti-actin (1:5000; Sigma-Aldrich).

    Techniques: Injection

    mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p < 0.05 and ** p < 0.01, vs. W, Tukey’s honestly significant difference (HSD). NS no significant difference compared with W, Tukey’s HSD. ( d – f ) Relative mRNA expression in primary RGCs. Primary RGCs isolated from 3-day-old rats by two-step immunopanning were cultured with or without KUS121 (50 µM) for 2 h and then with or without KUS121 and with or without NMDA (500 µM) for another 4 h. The relative expression levels of Edn1 ( d ), Ednrb ( e ) and Ednra ( f ) mRNA were analyzed using qRT-PCR. (-): without KUS121 without NMDA n = 3, C: with NMDA without KUS121, n = 3, K121: with NMDA with KUS121, n = 3, respectively. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. ** p < 0.01, control vs. KUS121, NS no significant difference compared with (-), Tukey’s HSD.

    Article Snippet: The primary antibodies used to probe the blots were mouse anti-EDN1 (1:400; Abcam, Cambridge, UK), rabbit anti-EDNRB (1:400; Alomone Labs, Jerusalem, Israel), rabbit anti-AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-AKT (1:2000, Cell Signaling Technology), rabbit anti-ERK (1:20,000, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-phospho-ERK (1:1000, Cell Signaling Technology) and anti-actin (1:5000; Sigma-Aldrich).

    Techniques: Expressing, Injection, Quantitative RT-PCR, Isolation, Cell Culture

    Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p < 0.01, vs. W, Tukey’s HSD. ( c – f ) Vertical sections of non-treated wild-type, NMDA-injected, NMDA-injected-KUS121-treated and NMDA-injected-KUS187-treated mice retinae were stained with an anti-EDN1 antibody or anti-EDNRB antibody. ( c , d ) Staining intensities of RGC layers with anti-EDN1 ( c ) or anti-EDNRB ( d ) antibody. The staining intensity of the RGC layer at distances of 400–800 μm from the optic nerve head was analyzed using BZ II Analyzer software. W: non-treated wild-type, C: NMDA-injected, K121: NMDA-injected-KUS121-treated, K187: NMDA-injected-KUS187-treated. ( n = 3, for C and K121, n = 2, for W and K187) NS no significant difference compared with W, Tukey’s HSD. ( e,f ) Vertical sections of non-treated wild-type (WT), NMDA-injected (control), NMDA-injected-KUS121-treated (K121) and NMDA-injected-KUS187-treated mice (K187) retinae. The black bar represents 100 µm. GCL ganglion cell layer, IPL inner plexiform layer.

    Article Snippet: The primary antibodies used to probe the blots were mouse anti-EDN1 (1:400; Abcam, Cambridge, UK), rabbit anti-EDNRB (1:400; Alomone Labs, Jerusalem, Israel), rabbit anti-AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-AKT (1:2000, Cell Signaling Technology), rabbit anti-ERK (1:20,000, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-phospho-ERK (1:1000, Cell Signaling Technology) and anti-actin (1:5000; Sigma-Aldrich).

    Techniques: Expressing, Labeling, Injection, Western Blot, Staining, Software

    EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Journal: Scientific Reports

    Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

    doi: 10.1038/s41598-022-20497-w

    Figure Lengend Snippet: EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p < 0.05, ** p < 0.01 and *** p < 0.001, compared with the control (in a and g ) or to the cells cultured in high glucose media (in h ), Tukey’s HSD.

    Article Snippet: The primary antibodies used to probe the blots were mouse anti-EDN1 (1:400; Abcam, Cambridge, UK), rabbit anti-EDNRB (1:400; Alomone Labs, Jerusalem, Israel), rabbit anti-AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-AKT (1:2000, Cell Signaling Technology), rabbit anti-ERK (1:20,000, Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-phospho-ERK (1:1000, Cell Signaling Technology) and anti-actin (1:5000; Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Labeling, Western Blot

    Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of endothelin-1 (ET-1) after ET B receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.

    Journal: PLoS ONE

    Article Title: Myocardial ischemia-reperfusion enhances transcriptional expression of endothelin-1 and vasoconstrictor ET B receptors via the protein kinase MEK-ERK1/2 signaling pathway in rat

    doi: 10.1371/journal.pone.0174119

    Figure Lengend Snippet: Concentration-response curves show the effect of increasing concentrations of Sarafotoxin 6c (S6c) in the left anterior descending artery (LAD) upstream of the ligature, LAD downstream of the ligature, and non-ischemic septal coronary artery (SCA) after treatment with (A) IR and vehicle or (B) U0126. The results are shown as mean values ± SEM (n = 6–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream. Concentration-response curves show the effect of increasing concentrations of endothelin-1 (ET-1) after ET B receptor desensitization and BQ788 treatment in the LAD upstream of the ligature, LAD downstream of the ligature, and non-ischemic SCA after treatment with (C) the vehicle or (D) U0126. The results are shown as mean values ± SEM (n = 4–7), *** P < 0.001; two-way ANOVA with repeated measurements; LAD downstream vs. upstream.

    Article Snippet: Membranes were subsequently blocked for 1 h in 2% ECL Prime Blocking Agent (GE Healthcare Life Sciences, Brøndby, Denmark) in TBS-T (TBS plus 0.1% Tween 20) before incubation overnight at 4°C on a rotor with the following primary antibodies diluted in TBS-T containing 0.02% sodium azide: polyclonal rabbit-anti-endothelin ET B receptor antibody (#AER-002, Alomone Laboratories, Israel 1:200); monoclonal mouse-anti-ET-1 antibody (#ab2786, Abcam, 1:1000); polyclonal rabbit-anti-endothelin ET A receptor antibody (#9780, Sigma, 1:1000); rabbit monoclonal anti-total-ERK antibody (#9101, Cell Signaling, 1:2000); or mouse monoclonal anti-pERK antibody (#9107, Cell Signaling, 1:1000).

    Techniques: Concentration Assay

    Antibodies.

    Journal: F1000Research

    Article Title: A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions

    doi: 10.12688/f1000research.7142.2

    Figure Lengend Snippet: Antibodies.

    Article Snippet: EDNRB Rabbit , Primary, indirect IF , 8μg/ml , Alomone labs #AER-002.

    Techniques:

    Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the ET B R mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative immunofluorescence staining on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).

    Journal: Open Forum Infectious Diseases

    Article Title: Human Cytomegalovirus Up-Regulates Endothelin Receptor Type B: Implication for Vasculopathies?

    doi: 10.1093/ofid/ofv155

    Figure Lengend Snippet: Effects of human cytomegalovirus (HCMV) on vascular endothelial and smooth muscle cells. The graphic depicts relative expression of endothelin receptor type B (ET B R) with indicated multiplicity of infection (moi) of HCMV as assayed by quantitative TaqMan polymerase chain reaction (PCR) at indicated days postinfection (dpi) for endothelial cells (human umbilical vein cells [HUVECs]; A.i) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs], A.ii) and human aortic SMCs [hSMCs], A.iii). The corresponding infection levels in HCMV-infected HUVECs (A.iv), hPASMCs (A.v) and hSMCs (A.vi), as assessed by quantitative TaqMan PCR are shown. The effects of UV-irradiated HCMV on the ET B R mRNA level in hPASMC (A.vii) and hSMC (A.viii) are illustrated. The uninfected cells at the indicated time course were used as a calibrator for relative quantification using the comparative 2 −ΔΔCt method with β 2 -microglobulin as an endogenous control (biological replicates for A.i-A.vi [n = 4] and A.vii and A.viii [n = 3] with technical duplicates for each PCR; values are mean ± standard deviation [SD]). Representative immunofluorescence staining on HCMV-infected endothelial cells (HUVECs; B.i) and SMCs (hPASMC, B.ii and hSMC, B.iii) with indicated MOI at 3 dpi. Arrows in B.i-B.iii depict HCMV-infected cells without up-regulation of ET B R. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI], blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). The corresponding quantitative results are shown for HCMV-infected HUVECs (B.iv), hPASMC (B.v) and hSMC (B.vi), by mean florescent intensity expressed as pixel sum as analyzed with Leica Application Suite Advanced Fluorescence software. At least 5 different areas of interest were quantified (values are mean ± SD; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001; n.s = not significant).

    Article Snippet: For dual staining, cells were incubated with rabbit anti-ET B R (1:200) (AER-002; Alomone Labs) for 30 minutes at room temperature and then washed and fixed with Reagent A followed by permeabilization with Reagent B (Molecular Probes; Invitrogen, Life Technologies) along with primary mouse anti-IE according to the manufacturer's protocol.

    Techniques: Expressing, Infection, Polymerase Chain Reaction, Irradiation, Standard Deviation, Immunofluorescence, Staining, Fluorescence, Software

    Human cytomegalovirus (HCMV) infection alters endothelin receptor type B (ET B R) cellular distribution, induces surface expression of ET B R and its effect on endothelin receptor type A (ET A R). (A) Representative immunofluorescence staining on HCMV-infected endothelial cells at 3 days postinfection to illustrate the subcellular distribution of ET B R in uninfected and infected cells. Arrow shows increased ET B R protein expression in the perinuclear region. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). (B) Representative immunofluorescence staining of ET B R and giantin, a Golgi marker (top panel), or PDI, an endoplasmic reticulum marker (bottom panel). Nucleus was stained with DAPI (blue). Flow cytometry analysis on the effects of HCMV-infected endothelial cells (human umbilical vein cells [HUVECs]) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs] and human aortic SMCs [hSMCs]) on ET B R at indicated days postinfection (dpi). (C) The effects of different multiplicity of infection (moi) of HCMV on ET B R surface expression at 1, 3, and 5 dpi (left panel) in HUVECs. The HCMV IE is shown on the right panel. Y-axis denotes log intensity of ET B R (left panel) or IE (right panel), whereas x-axis is side scatter in linear scale (SS Lin). (D) The effects of ultraviolet (UV)-irradiated HCMV on ET B R surface expression in HUVECs. Little or no IE was detected with the UV treatment (bottom row). (E) The effects of UV-irradiated HCMV (middle row) or HCMV (bottom row) on ET B R surface expression in SMCs, hPASMCs (left column) and hSMCs (right column) at 5 dpi. Y-axis denotes log intensity of ET B R (left panel), whereas x-axis is SS Lin. The effects of HCMV on ET A R mRNA level in smooth muscle cells, hPASMC (F) and hSMC (G) (MOI; biological replicates, n = 3 with technical duplicates for each polymerase chain reaction; values are mean ± standard deviation; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001).

    Journal: Open Forum Infectious Diseases

    Article Title: Human Cytomegalovirus Up-Regulates Endothelin Receptor Type B: Implication for Vasculopathies?

    doi: 10.1093/ofid/ofv155

    Figure Lengend Snippet: Human cytomegalovirus (HCMV) infection alters endothelin receptor type B (ET B R) cellular distribution, induces surface expression of ET B R and its effect on endothelin receptor type A (ET A R). (A) Representative immunofluorescence staining on HCMV-infected endothelial cells at 3 days postinfection to illustrate the subcellular distribution of ET B R in uninfected and infected cells. Arrow shows increased ET B R protein expression in the perinuclear region. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of ET B R or HCMV immediate-early (IE) was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively (biological replicates, n = 3). (B) Representative immunofluorescence staining of ET B R and giantin, a Golgi marker (top panel), or PDI, an endoplasmic reticulum marker (bottom panel). Nucleus was stained with DAPI (blue). Flow cytometry analysis on the effects of HCMV-infected endothelial cells (human umbilical vein cells [HUVECs]) or smooth muscle cells (SMCs) (human pulmonary arterial SMCs [hPASMCs] and human aortic SMCs [hSMCs]) on ET B R at indicated days postinfection (dpi). (C) The effects of different multiplicity of infection (moi) of HCMV on ET B R surface expression at 1, 3, and 5 dpi (left panel) in HUVECs. The HCMV IE is shown on the right panel. Y-axis denotes log intensity of ET B R (left panel) or IE (right panel), whereas x-axis is side scatter in linear scale (SS Lin). (D) The effects of ultraviolet (UV)-irradiated HCMV on ET B R surface expression in HUVECs. Little or no IE was detected with the UV treatment (bottom row). (E) The effects of UV-irradiated HCMV (middle row) or HCMV (bottom row) on ET B R surface expression in SMCs, hPASMCs (left column) and hSMCs (right column) at 5 dpi. Y-axis denotes log intensity of ET B R (left panel), whereas x-axis is SS Lin. The effects of HCMV on ET A R mRNA level in smooth muscle cells, hPASMC (F) and hSMC (G) (MOI; biological replicates, n = 3 with technical duplicates for each polymerase chain reaction; values are mean ± standard deviation; P < .05 was considered to be statistically significant; * P = .01–.05; ** P = .001–.01; *** P < .001).

    Article Snippet: For dual staining, cells were incubated with rabbit anti-ET B R (1:200) (AER-002; Alomone Labs) for 30 minutes at room temperature and then washed and fixed with Reagent A followed by permeabilization with Reagent B (Molecular Probes; Invitrogen, Life Technologies) along with primary mouse anti-IE according to the manufacturer's protocol.

    Techniques: Infection, Expressing, Immunofluorescence, Staining, Marker, Flow Cytometry, Irradiation, Polymerase Chain Reaction, Standard Deviation

    Human cytomegalovirus (HCMV) antigens are closely associated with endothelin receptor type B (ET B R) in human carotid atherosclerotic plaques. Representative photomicrographs of immunohistochemistry and immunofluorescence (IF) staining on frozen human carotid atherosclerotic plaques. (A) Immunohistochemistry staining to show colocalization of HCMV immediate-early (IE) and ET B R in Von Willebrand factor (vWF)-positive cells as revealed by chromogen diaminobenzidine, brown products in a boxed area with consecutive sections (left column; right column is a higher magnification of the boxed areas). (B) The IF staining on a human carotid atherosclerotic plaque to illustrate the colocalization of HCMV-IE with ET B R in fibroblast-like cells. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of HCMV-IE or ET B R was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively. Twenty human carotid atherosclerotic plaques were examined.

    Journal: Open Forum Infectious Diseases

    Article Title: Human Cytomegalovirus Up-Regulates Endothelin Receptor Type B: Implication for Vasculopathies?

    doi: 10.1093/ofid/ofv155

    Figure Lengend Snippet: Human cytomegalovirus (HCMV) antigens are closely associated with endothelin receptor type B (ET B R) in human carotid atherosclerotic plaques. Representative photomicrographs of immunohistochemistry and immunofluorescence (IF) staining on frozen human carotid atherosclerotic plaques. (A) Immunohistochemistry staining to show colocalization of HCMV immediate-early (IE) and ET B R in Von Willebrand factor (vWF)-positive cells as revealed by chromogen diaminobenzidine, brown products in a boxed area with consecutive sections (left column; right column is a higher magnification of the boxed areas). (B) The IF staining on a human carotid atherosclerotic plaque to illustrate the colocalization of HCMV-IE with ET B R in fibroblast-like cells. Nucleus was stained with 4′-6-diamidino-2-phenylindole ([DAPI] blue), and the immunoreactivity of HCMV-IE or ET B R was revealed with Alexa Fluor 594 (red) or Alexa Fluor 488 (green), respectively. Twenty human carotid atherosclerotic plaques were examined.

    Article Snippet: For dual staining, cells were incubated with rabbit anti-ET B R (1:200) (AER-002; Alomone Labs) for 30 minutes at room temperature and then washed and fixed with Reagent A followed by permeabilization with Reagent B (Molecular Probes; Invitrogen, Life Technologies) along with primary mouse anti-IE according to the manufacturer's protocol.

    Techniques: Immunohistochemistry, Immunofluorescence, Staining