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calcium ionophore a23187 io  (Tocris)
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Tocris calcium ionophore a23187 io
AngII induces CN-dependent gene and protein expression of Rcan1 in VSMCs. (A–D) VSMCs were stimulated with 1 µM AngII after pretreatment for 1 h with 200 ng/ml CsA, 1 µM of the AT 1 blocker ZD7155, or 10 µM of the AT 2 blocker PD123319. (A) RNA expression in VSMCs stimulated with AngII for 4 h after pretreatment or not with CsA was analyzed by hybridization of arrays. Expression of the indicated genes in the array under each experimental condition is shown as fold changes (log 2 ) relative to control cells (P < 0.05 vs. control). (B) qPCR analysis of Rcan1 mRNA expression after AngII treatment for 1 and 4 h. mRNA amounts were normalized to Tbp expression and data are expressed as the fold change relative to nonstimulated cells. Means of triplicate qPCR determinations for each condition are shown from a representative experiment of five performed. (C) Time course Rcan1 immunoblot in VSMCs treated as indicated. (D) Immunoblots showing the effect of AT 1 or AT 2 blockers on AngII-stimulated Rcan1-4 expression. PIo, cells stimulated with a phorbol ester plus calcium ionophore <t>A23187.</t> (E) Immunoblots showing Rcan1-4 expression in VSMCs infected with lentiviruses expressing GFP-tagged LxVP or LxVPmutant peptides and treated as indicated. (C–E) Tubulin expression was used as loading control. Representative experiments of four to six performed are shown. (F) Relative luciferase activity from VSMCs transfected with the indicated reporters and treated as indicated above. PIo treatment was used as a positive control. Values are means ± SEM of triplicate luciferase determinations for each condition from five experiments.
Calcium Ionophore A23187 Io, supplied by Tocris, 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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AngII induces CN-dependent gene and protein expression of Rcan1 in VSMCs. (A–D) VSMCs were stimulated with 1 µM AngII after pretreatment for 1 h with 200 ng/ml CsA, 1 µM of the AT 1 blocker ZD7155, or 10 µM of the AT 2 blocker PD123319. (A) RNA expression in VSMCs stimulated with AngII for 4 h after pretreatment or not with CsA was analyzed by hybridization of arrays. Expression of the indicated genes in the array under each experimental condition is shown as fold changes (log 2 ) relative to control cells (P < 0.05 vs. control). (B) qPCR analysis of Rcan1 mRNA expression after AngII treatment for 1 and 4 h. mRNA amounts were normalized to Tbp expression and data are expressed as the fold change relative to nonstimulated cells. Means of triplicate qPCR determinations for each condition are shown from a representative experiment of five performed. (C) Time course Rcan1 immunoblot in VSMCs treated as indicated. (D) Immunoblots showing the effect of AT 1 or AT 2 blockers on AngII-stimulated Rcan1-4 expression. PIo, cells stimulated with a phorbol ester plus calcium ionophore A23187. (E) Immunoblots showing Rcan1-4 expression in VSMCs infected with lentiviruses expressing GFP-tagged LxVP or LxVPmutant peptides and treated as indicated. (C–E) Tubulin expression was used as loading control. Representative experiments of four to six performed are shown. (F) Relative luciferase activity from VSMCs transfected with the indicated reporters and treated as indicated above. PIo treatment was used as a positive control. Values are means ± SEM of triplicate luciferase determinations for each condition from five experiments.

Journal: The Journal of Experimental Medicine

Article Title: Regulator of calcineurin 1 mediates pathological vascular wall remodeling

doi: 10.1084/jem.20110503

Figure Lengend Snippet: AngII induces CN-dependent gene and protein expression of Rcan1 in VSMCs. (A–D) VSMCs were stimulated with 1 µM AngII after pretreatment for 1 h with 200 ng/ml CsA, 1 µM of the AT 1 blocker ZD7155, or 10 µM of the AT 2 blocker PD123319. (A) RNA expression in VSMCs stimulated with AngII for 4 h after pretreatment or not with CsA was analyzed by hybridization of arrays. Expression of the indicated genes in the array under each experimental condition is shown as fold changes (log 2 ) relative to control cells (P < 0.05 vs. control). (B) qPCR analysis of Rcan1 mRNA expression after AngII treatment for 1 and 4 h. mRNA amounts were normalized to Tbp expression and data are expressed as the fold change relative to nonstimulated cells. Means of triplicate qPCR determinations for each condition are shown from a representative experiment of five performed. (C) Time course Rcan1 immunoblot in VSMCs treated as indicated. (D) Immunoblots showing the effect of AT 1 or AT 2 blockers on AngII-stimulated Rcan1-4 expression. PIo, cells stimulated with a phorbol ester plus calcium ionophore A23187. (E) Immunoblots showing Rcan1-4 expression in VSMCs infected with lentiviruses expressing GFP-tagged LxVP or LxVPmutant peptides and treated as indicated. (C–E) Tubulin expression was used as loading control. Representative experiments of four to six performed are shown. (F) Relative luciferase activity from VSMCs transfected with the indicated reporters and treated as indicated above. PIo treatment was used as a positive control. Values are means ± SEM of triplicate luciferase determinations for each condition from five experiments.

Article Snippet: 1 μM calcium ionophore A23187 (Io), 10 −6 M AT1RB (ZD-7155 hydrochloride), and 200 ng/ml CsA were obtained from EMD, Tocris Bioscience, and LD laboratories, respectively.

Techniques: Expressing, RNA Expression, Hybridization, Control, Western Blot, Infection, Luciferase, Activity Assay, Transfection, Positive Control