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Fig. 7. mAChR stimulation induces STIM1–Orai1 and <t>TRPC1–Orai1</t> interactions. (A) Cells were pre-treated with either vehicle or 100 nM atropine (atrop) for 5 min and then exposed to vehicle or 10 μM CCh for 5 min. Cell lysates were incubated with anti-Orai1 antibody and STIM1 and Orai1 immunoreactivities were analyzed in immunoprecipitates and cell lysates. STIM1 immunoreactivity in immunoprecipitates (IP) was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of four experiments. *P b 0.05 vs control (vehicle), #P b 0.05 by ANOVA followed by Newman–Keuls post hoc test. (B) Cells were treated with either vehicle or CCh as described in (A) and cell lysates were subjected to immunoprecipitation with pre-immune IgG. Immunoprecipitates and cell lysates were then analyzed for STIM1 and Orai1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments. (C) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-Orai1 antibody. Immunoprecipitates and cell lysates were analyzed for TRPC1 and Orai1 immunoreactivities. The amount of TRPC1 in immunoprecipitates was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of three experiments. *P b 0.05 vs control (vehicle). (D) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-TRPC1 antibody. Immunoprecipitates and cell lysates were analyzed for STIM1 and TRPC1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments and is representative of a total of four experiments.
Anti Transient Receptor Potential Canonical 1 Trpc1, supplied by Santa Cruz Biotechnology, 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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Fig. 7. mAChR stimulation induces STIM1–Orai1 and <t>TRPC1–Orai1</t> interactions. (A) Cells were pre-treated with either vehicle or 100 nM atropine (atrop) for 5 min and then exposed to vehicle or 10 μM CCh for 5 min. Cell lysates were incubated with anti-Orai1 antibody and STIM1 and Orai1 immunoreactivities were analyzed in immunoprecipitates and cell lysates. STIM1 immunoreactivity in immunoprecipitates (IP) was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of four experiments. *P b 0.05 vs control (vehicle), #P b 0.05 by ANOVA followed by Newman–Keuls post hoc test. (B) Cells were treated with either vehicle or CCh as described in (A) and cell lysates were subjected to immunoprecipitation with pre-immune IgG. Immunoprecipitates and cell lysates were then analyzed for STIM1 and Orai1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments. (C) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-Orai1 antibody. Immunoprecipitates and cell lysates were analyzed for TRPC1 and Orai1 immunoreactivities. The amount of TRPC1 in immunoprecipitates was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of three experiments. *P b 0.05 vs control (vehicle). (D) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-TRPC1 antibody. Immunoprecipitates and cell lysates were analyzed for STIM1 and TRPC1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments and is representative of a total of four experiments.
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N/A
The protein encoded by this gene is a membrane protein that can form a non selective channel permeable to calcium and other cations The encoded protein appears to be induced to form channels by a
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Fig. 7. mAChR stimulation induces STIM1–Orai1 and TRPC1–Orai1 interactions. (A) Cells were pre-treated with either vehicle or 100 nM atropine (atrop) for 5 min and then exposed to vehicle or 10 μM CCh for 5 min. Cell lysates were incubated with anti-Orai1 antibody and STIM1 and Orai1 immunoreactivities were analyzed in immunoprecipitates and cell lysates. STIM1 immunoreactivity in immunoprecipitates (IP) was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of four experiments. *P b 0.05 vs control (vehicle), #P b 0.05 by ANOVA followed by Newman–Keuls post hoc test. (B) Cells were treated with either vehicle or CCh as described in (A) and cell lysates were subjected to immunoprecipitation with pre-immune IgG. Immunoprecipitates and cell lysates were then analyzed for STIM1 and Orai1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments. (C) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-Orai1 antibody. Immunoprecipitates and cell lysates were analyzed for TRPC1 and Orai1 immunoreactivities. The amount of TRPC1 in immunoprecipitates was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of three experiments. *P b 0.05 vs control (vehicle). (D) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-TRPC1 antibody. Immunoprecipitates and cell lysates were analyzed for STIM1 and TRPC1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments and is representative of a total of four experiments.

Journal: Biochimica et biophysica acta

Article Title: Involvement of store-operated Ca(2+) entry in activation of AMP-activated protein kinase and stimulation of glucose uptake by M3 muscarinic acetylcholine receptors in human neuroblastoma cells.

doi: 10.1016/j.bbamcr.2014.09.012

Figure Lengend Snippet: Fig. 7. mAChR stimulation induces STIM1–Orai1 and TRPC1–Orai1 interactions. (A) Cells were pre-treated with either vehicle or 100 nM atropine (atrop) for 5 min and then exposed to vehicle or 10 μM CCh for 5 min. Cell lysates were incubated with anti-Orai1 antibody and STIM1 and Orai1 immunoreactivities were analyzed in immunoprecipitates and cell lysates. STIM1 immunoreactivity in immunoprecipitates (IP) was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of four experiments. *P b 0.05 vs control (vehicle), #P b 0.05 by ANOVA followed by Newman–Keuls post hoc test. (B) Cells were treated with either vehicle or CCh as described in (A) and cell lysates were subjected to immunoprecipitation with pre-immune IgG. Immunoprecipitates and cell lysates were then analyzed for STIM1 and Orai1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments. (C) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-Orai1 antibody. Immunoprecipitates and cell lysates were analyzed for TRPC1 and Orai1 immunoreactivities. The amount of TRPC1 in immunoprecipitates was normalized to that present in cell lysates. Values are expressed as percent of control and are the mean ± SEM of three experiments. *P b 0.05 vs control (vehicle). (D) Cells were treated with either vehicle or CCh as in (A) and cell lysates were immunoprecipitated with anti-TRPC1 antibody. Immunoprecipitates and cell lysates were analyzed for STIM1 and TRPC1 immunoreactivities. The image shows immunoblots of sample pair (vehicle and CCh) from two independent experiments and is representative of a total of four experiments.

Article Snippet: The supernatants (~700 μg of protein) were incubated overnight at 4 °C with either antiOrai1 (3 μg), anti-transient receptor potential canonical 1 (TRPC1) (2 μg) antibodies or pre-immune IgG (3 μg) (Santa Cruz Biotechnology).

Techniques: Incubation, Control, Immunoprecipitation, Western Blot