trpc3  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    Alomone Labs trpc3
    <t>TRPC3</t> forms a stable ternary complex with Nox2 and p22 phox . ( a , b ) Expression of Nox2 and p22 phox proteins in HEK293 cells that express a different combination of TRPC3-GFP and GFP. Results of a quantitative analysis are shown in ( b ) (n = 3). ( c ) Nox2 mRNA amounts in HEK293 cells co-expressing Nox2 with GFP or TRPC3-GFP (n = 3). ( d ) Increased Nox2 and p22 phox protein in HEK293 cells co-expressing pore-dead mutant of TRPC3 (n = 3). ( e ) Interaction of TRPC3 with Nox2 in HEK293 cells. Immunoprecipitation was performed using an anti-flag antibody. ( f ) Nox2 protein expression in HEK293 cells expressing TRPC3 alone or co-expressing TRPC3 and TRPC6 (n = 3). Error bars, s.e.m. *P
    Trpc3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trpc3/product/Alomone Labs
    Average 95 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    trpc3 - by Bioz Stars, 2022-08
    95/100 stars

    Images

    1) Product Images from "TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling"

    Article Title: TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling

    Journal: Scientific Reports

    doi: 10.1038/srep37001

    TRPC3 forms a stable ternary complex with Nox2 and p22 phox . ( a , b ) Expression of Nox2 and p22 phox proteins in HEK293 cells that express a different combination of TRPC3-GFP and GFP. Results of a quantitative analysis are shown in ( b ) (n = 3). ( c ) Nox2 mRNA amounts in HEK293 cells co-expressing Nox2 with GFP or TRPC3-GFP (n = 3). ( d ) Increased Nox2 and p22 phox protein in HEK293 cells co-expressing pore-dead mutant of TRPC3 (n = 3). ( e ) Interaction of TRPC3 with Nox2 in HEK293 cells. Immunoprecipitation was performed using an anti-flag antibody. ( f ) Nox2 protein expression in HEK293 cells expressing TRPC3 alone or co-expressing TRPC3 and TRPC6 (n = 3). Error bars, s.e.m. *P
    Figure Legend Snippet: TRPC3 forms a stable ternary complex with Nox2 and p22 phox . ( a , b ) Expression of Nox2 and p22 phox proteins in HEK293 cells that express a different combination of TRPC3-GFP and GFP. Results of a quantitative analysis are shown in ( b ) (n = 3). ( c ) Nox2 mRNA amounts in HEK293 cells co-expressing Nox2 with GFP or TRPC3-GFP (n = 3). ( d ) Increased Nox2 and p22 phox protein in HEK293 cells co-expressing pore-dead mutant of TRPC3 (n = 3). ( e ) Interaction of TRPC3 with Nox2 in HEK293 cells. Immunoprecipitation was performed using an anti-flag antibody. ( f ) Nox2 protein expression in HEK293 cells expressing TRPC3 alone or co-expressing TRPC3 and TRPC6 (n = 3). Error bars, s.e.m. *P

    Techniques Used: Expressing, Mutagenesis, Immunoprecipitation

    TRPC3 plays a critical role in Mechanical stretch-induced ROS production. ( a , b ) Effects of siRNA targeting TRPC1, C3 or C6 on mechanical stretch (MS)-induced ROS production (n = 3). ( c ) mRNA expression of either TRPC1 or TRPC6 in NRCM transfected with siRNAs against either TRPC1 or TRPC6, respectively (n = 3). ( d,e ) Time courses of MS-induced ROS production in NRCMs treated with GsMTx-4 (1 μM; ( d ) or TRPV4 inhibitor (RN1734, 50 μM; ( e ) Reagents were added to cells 5 min before MS (n = 3). ( f ) MS-induced ROS production in TRPC(1–7)-deficient MEF cells expressing TRPC3, TRPC6, TRPC7, or LacZ (n = 30). Data are representative of three independent experiments. ( g–i ) Effect of TRPC3 siRNA on the protein abundances of TRPC3 ( h ) and Nox2 ( i ) protein expressions in NRCMs (n = 3). Error bars, s.e.m. *P
    Figure Legend Snippet: TRPC3 plays a critical role in Mechanical stretch-induced ROS production. ( a , b ) Effects of siRNA targeting TRPC1, C3 or C6 on mechanical stretch (MS)-induced ROS production (n = 3). ( c ) mRNA expression of either TRPC1 or TRPC6 in NRCM transfected with siRNAs against either TRPC1 or TRPC6, respectively (n = 3). ( d,e ) Time courses of MS-induced ROS production in NRCMs treated with GsMTx-4 (1 μM; ( d ) or TRPV4 inhibitor (RN1734, 50 μM; ( e ) Reagents were added to cells 5 min before MS (n = 3). ( f ) MS-induced ROS production in TRPC(1–7)-deficient MEF cells expressing TRPC3, TRPC6, TRPC7, or LacZ (n = 30). Data are representative of three independent experiments. ( g–i ) Effect of TRPC3 siRNA on the protein abundances of TRPC3 ( h ) and Nox2 ( i ) protein expressions in NRCMs (n = 3). Error bars, s.e.m. *P

    Techniques Used: Mass Spectrometry, Expressing, Transfection

    TRPC3 prevents Nox2 protein from proteasomal degradation. ( a–e ) Abundances of Nox2 protein ( a , b ) and mRNAs of TRPC3 ( c ), Nox2 ( d ), and p22 phox ( e ) in NRCM transfected with siRNAs targeting TRPC3 with or without MG132. Cells were treated with siRNAs and MG132 (1 μM) simultaneously (n = 3). ( f , g ) Effect of siRNA targeting TRPC3 on Nox2 protein abundance in cell surface (Surface) and total lysates (Total) from NRCMs (n = 3). GAPDH was used as an internal control. Error bars, s.e.m. *P
    Figure Legend Snippet: TRPC3 prevents Nox2 protein from proteasomal degradation. ( a–e ) Abundances of Nox2 protein ( a , b ) and mRNAs of TRPC3 ( c ), Nox2 ( d ), and p22 phox ( e ) in NRCM transfected with siRNAs targeting TRPC3 with or without MG132. Cells were treated with siRNAs and MG132 (1 μM) simultaneously (n = 3). ( f , g ) Effect of siRNA targeting TRPC3 on Nox2 protein abundance in cell surface (Surface) and total lysates (Total) from NRCMs (n = 3). GAPDH was used as an internal control. Error bars, s.e.m. *P

    Techniques Used: Transfection

    TRPC3 forms a stable ternary complex with Nox2 and p22 phox proteins in endogenously p22 phox -absent CHO cells. ( a ) Expression of Nox2 and p22 phox proteins in CHO cells that express a different combination of TRPC3-GFP and GFP. ( b ) Results of quantitative analysis (n = 3). ( c ) Expression of Nox2 and p22 phox co-expressed with either GFP or TRPC3-GFP in MG132 (10 μM)-treated CHO cells. ( d ) Graphs depict the relative expression of either Nox2 or p22 phox protein to that in non-treated cells. Band intensities were normalized by GAPDH. ( e–g ) Interaction of TRPC3 with p22 phox and Nox2 in CHO cells. ( h ) Localization of Nox2 in CHO cells co-expressing Nox2 with TRPC3-GFP (or GFP-F). Error bars, s.e.m. *P
    Figure Legend Snippet: TRPC3 forms a stable ternary complex with Nox2 and p22 phox proteins in endogenously p22 phox -absent CHO cells. ( a ) Expression of Nox2 and p22 phox proteins in CHO cells that express a different combination of TRPC3-GFP and GFP. ( b ) Results of quantitative analysis (n = 3). ( c ) Expression of Nox2 and p22 phox co-expressed with either GFP or TRPC3-GFP in MG132 (10 μM)-treated CHO cells. ( d ) Graphs depict the relative expression of either Nox2 or p22 phox protein to that in non-treated cells. Band intensities were normalized by GAPDH. ( e–g ) Interaction of TRPC3 with p22 phox and Nox2 in CHO cells. ( h ) Localization of Nox2 in CHO cells co-expressing Nox2 with TRPC3-GFP (or GFP-F). Error bars, s.e.m. *P

    Techniques Used: Expressing

    Formation of a TRPC3/Nox2 complex promotes TRPC3 channel activity through stabilization at the plasma membrane. ( a ) Effect of Nox2 siRNA on expression of TRPC3 in NRCMs (n = 3). ( b ) Representative images showing the levels of TRPC3-GFP and GFP expression in HEK293 cells co-expressing p22 phox or Nox2 (n = 3). ( c ) Expression of TRPC3-GFP mRNA in HEK293 cells co-expressing p22 phox or Nox2 (n = 3). ( d–f ) Representative time courses of TRPC3 currents ( d ) and the current-voltage (I-V) relationships ( e ) and peak TRPC3 current densities at −60 mV ( f ) induced by 100 μM carbachol (CCh) in HEK293 cells expressing TRPC3-mCherry alone or with p22 phox , Nox2, both p22 phox and Nox2, or Nox2 treated with DPI. DPI (0.3 μM) was treated 1 min before CCh stimulation. ( g , h ) Representative Ca 2+ responses in the presence ( g ) or absence ( h ) of pyrazole-3 (Pyr3, 1 μM) upon mechanical stretch (MS) application. ( i ) Peak Ca 2+ increases after MS in NRCMs treated with (n = 61) or without Pyr3 (n = 78). ( j ) Changes of minimal [Ca 2+ ] i before and after MS application. Minimal [Ca 2+ ] i from Ca 2+ responses in every 1 min were analyzed and represented as diastolic [Ca 2+ ] i . ( k ) Schematic images showing phosphorylation of p47 phox via TRPC3-PKCβ activation induced by MS in the heart. ( l–n ) Effects of TRPC3 ( l , m ) or PKCβ ( n) ; 10 μM Gö6976) inhibitors on p47 phox phosphorylation induced by MS in NRCMs (n = 3). ( o ) MS-induced ROS generation in NRCMs treated with a PKCβ inhibitor (n = 3). ( p ) Co-immunoprecipitation of TRPC3 with PKCβ, Nox2 and p22 phox in mouse hearts 1week after TAC operation (n = 3). Error bars, s.e.m.*P
    Figure Legend Snippet: Formation of a TRPC3/Nox2 complex promotes TRPC3 channel activity through stabilization at the plasma membrane. ( a ) Effect of Nox2 siRNA on expression of TRPC3 in NRCMs (n = 3). ( b ) Representative images showing the levels of TRPC3-GFP and GFP expression in HEK293 cells co-expressing p22 phox or Nox2 (n = 3). ( c ) Expression of TRPC3-GFP mRNA in HEK293 cells co-expressing p22 phox or Nox2 (n = 3). ( d–f ) Representative time courses of TRPC3 currents ( d ) and the current-voltage (I-V) relationships ( e ) and peak TRPC3 current densities at −60 mV ( f ) induced by 100 μM carbachol (CCh) in HEK293 cells expressing TRPC3-mCherry alone or with p22 phox , Nox2, both p22 phox and Nox2, or Nox2 treated with DPI. DPI (0.3 μM) was treated 1 min before CCh stimulation. ( g , h ) Representative Ca 2+ responses in the presence ( g ) or absence ( h ) of pyrazole-3 (Pyr3, 1 μM) upon mechanical stretch (MS) application. ( i ) Peak Ca 2+ increases after MS in NRCMs treated with (n = 61) or without Pyr3 (n = 78). ( j ) Changes of minimal [Ca 2+ ] i before and after MS application. Minimal [Ca 2+ ] i from Ca 2+ responses in every 1 min were analyzed and represented as diastolic [Ca 2+ ] i . ( k ) Schematic images showing phosphorylation of p47 phox via TRPC3-PKCβ activation induced by MS in the heart. ( l–n ) Effects of TRPC3 ( l , m ) or PKCβ ( n) ; 10 μM Gö6976) inhibitors on p47 phox phosphorylation induced by MS in NRCMs (n = 3). ( o ) MS-induced ROS generation in NRCMs treated with a PKCβ inhibitor (n = 3). ( p ) Co-immunoprecipitation of TRPC3 with PKCβ, Nox2 and p22 phox in mouse hearts 1week after TAC operation (n = 3). Error bars, s.e.m.*P

    Techniques Used: Activity Assay, Expressing, Mass Spectrometry, Activation Assay, Immunoprecipitation

    TRPC3 deletion suppresses TAC-induced LV dysfunction and dilation through Nox2 inhibition. ( a ) Left ventricular end-diastolic pressure (LVEDP; left) and dP/dT max (right) in TAC-operated TRPC3 (+/+) (n = 13) and TRPC3 (−/−) (n = 12) mice 6 week post-operation. ( b ) Myocardial malondialdehyde concentrations 1 week after TAC (n = 4). ( c ) Abundance of Nox2 protein in TRPC3 (+/+) and TRPC3 (−/−) hearts 1 week after TAC (n = 3). ( d ) Representative immunofluorescence images of TRPC3, p22 phox , and caveolin-3 (Cav-3) in adult mouse cardiomyocytes isolated from muscle LIM protein-deficient hearts. ( e ) Representative immunofluorescence images of p22 phox in adult mouse cardiomyocytes: green, anti-p22 phox ; blue, DAPI. ( f ) Relative abundances of p22 phox and Nox2 mRNA in mouse hearts 1 week after TAC (n = 4). ( g ) Abundance of Nox2 protein in TRPC6 (+/+) and TRPC6 (−/−) hearts 1 week after TAC (n = 3). Error bars, s.e.m. *P
    Figure Legend Snippet: TRPC3 deletion suppresses TAC-induced LV dysfunction and dilation through Nox2 inhibition. ( a ) Left ventricular end-diastolic pressure (LVEDP; left) and dP/dT max (right) in TAC-operated TRPC3 (+/+) (n = 13) and TRPC3 (−/−) (n = 12) mice 6 week post-operation. ( b ) Myocardial malondialdehyde concentrations 1 week after TAC (n = 4). ( c ) Abundance of Nox2 protein in TRPC3 (+/+) and TRPC3 (−/−) hearts 1 week after TAC (n = 3). ( d ) Representative immunofluorescence images of TRPC3, p22 phox , and caveolin-3 (Cav-3) in adult mouse cardiomyocytes isolated from muscle LIM protein-deficient hearts. ( e ) Representative immunofluorescence images of p22 phox in adult mouse cardiomyocytes: green, anti-p22 phox ; blue, DAPI. ( f ) Relative abundances of p22 phox and Nox2 mRNA in mouse hearts 1 week after TAC (n = 4). ( g ) Abundance of Nox2 protein in TRPC6 (+/+) and TRPC6 (−/−) hearts 1 week after TAC (n = 3). Error bars, s.e.m. *P

    Techniques Used: Inhibition, Mouse Assay, Immunofluorescence, Isolation

    Physical interaction between TRPC3 and Nox2 is critical for stabilization of Nox2. ( a ) Schematic illustration of TRPC3 terminal deletion mutants. ( b , c ) Expression of Nox2 and p22 phox co-expressed with TRPC3 deletion mutants in HEK293 cells (n = 3). ( d ) OAG-induced ROS production in NRCMs expressing Nox2-interacting TRPC3 C-terminal fragment (C3-C fragment) (n = 20–28). ( e ) Co-immunoprecipitation of TRPC3 with Nox2 in the presence or absence of C3-C fragment. Representative blot from three independent experiments was shown. ( f ) ATP (100 μM)-induced Ca 2+ responses in HEK293 cells expressing TRPC3 with or without C3-C fragment (n = 35–51). Timing of solution exchanges were indicated by horizontal bars above the graph. ( g ) Model of the regulation of TRPC3-Nox2 stability and induction of LV dysfunction induced by diastolic stretch of cardiomyocytes. Error bars, s.e.m.*P
    Figure Legend Snippet: Physical interaction between TRPC3 and Nox2 is critical for stabilization of Nox2. ( a ) Schematic illustration of TRPC3 terminal deletion mutants. ( b , c ) Expression of Nox2 and p22 phox co-expressed with TRPC3 deletion mutants in HEK293 cells (n = 3). ( d ) OAG-induced ROS production in NRCMs expressing Nox2-interacting TRPC3 C-terminal fragment (C3-C fragment) (n = 20–28). ( e ) Co-immunoprecipitation of TRPC3 with Nox2 in the presence or absence of C3-C fragment. Representative blot from three independent experiments was shown. ( f ) ATP (100 μM)-induced Ca 2+ responses in HEK293 cells expressing TRPC3 with or without C3-C fragment (n = 35–51). Timing of solution exchanges were indicated by horizontal bars above the graph. ( g ) Model of the regulation of TRPC3-Nox2 stability and induction of LV dysfunction induced by diastolic stretch of cardiomyocytes. Error bars, s.e.m.*P

    Techniques Used: Expressing, Immunoprecipitation

    2) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    3) Product Images from "Overexpression of TRPC3 increases apoptosis but not necrosis in response to ischemia/reperfusion in adult mouse cardiomyocytes"

    Article Title: Overexpression of TRPC3 increases apoptosis but not necrosis in response to ischemia/reperfusion in adult mouse cardiomyocytes

    Journal: American journal of physiology. Cell physiology

    doi: 10.1152/ajpcell.00313.2007

    After 1h of isolation, cardiomyocytes from WT and TRPC3 transgenic mice were treated with 10 ng/ml TNF-α for 2h or 18h, viability and apoptosis were measured by Annexin V-propidium iodide staining. Data presented as mean ± SEM of six individual experiments, i.e., six separate mouse cardiomyocytes isolations; with at least 300 cells counted per experiment under each condition. * = p
    Figure Legend Snippet: After 1h of isolation, cardiomyocytes from WT and TRPC3 transgenic mice were treated with 10 ng/ml TNF-α for 2h or 18h, viability and apoptosis were measured by Annexin V-propidium iodide staining. Data presented as mean ± SEM of six individual experiments, i.e., six separate mouse cardiomyocytes isolations; with at least 300 cells counted per experiment under each condition. * = p

    Techniques Used: Isolation, Transgenic Assay, Mouse Assay, Staining

    A) Bright field phase contrast images of cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice at baseline, at the end of 90 min ischemia and after 90 min ischemia and 3 hours reperfusion; B) cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice after 90 min ischemia and 3 hours reperfusion showing Annexin V and propidium iodide staining. Arrows in merged image indicate necrotic cells, staining positive for both Annexin V and propidium iodide.
    Figure Legend Snippet: A) Bright field phase contrast images of cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice at baseline, at the end of 90 min ischemia and after 90 min ischemia and 3 hours reperfusion; B) cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice after 90 min ischemia and 3 hours reperfusion showing Annexin V and propidium iodide staining. Arrows in merged image indicate necrotic cells, staining positive for both Annexin V and propidium iodide.

    Techniques Used: Transgenic Assay, Mouse Assay, Staining

    A) Cell viability assessed by % rod shaped cells; B) % apoptotic cells indicated by Annexin V positive and propidium iodide negative staining; C) % necrotic cells indicated by Annexin V and propidium iodide positive staining in cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice at baseline, at the end of 90 min ischemia and after 90 min ischemia and 3 hours reperfusion (I/R). Data presented as mean ± SEM of six individual experiments (i.e., six separate mouse cardiomyocytes isolations; with at least 300 cells counted per experiment under each condition). # = p
    Figure Legend Snippet: A) Cell viability assessed by % rod shaped cells; B) % apoptotic cells indicated by Annexin V positive and propidium iodide negative staining; C) % necrotic cells indicated by Annexin V and propidium iodide positive staining in cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice at baseline, at the end of 90 min ischemia and after 90 min ischemia and 3 hours reperfusion (I/R). Data presented as mean ± SEM of six individual experiments (i.e., six separate mouse cardiomyocytes isolations; with at least 300 cells counted per experiment under each condition). # = p

    Techniques Used: Negative Staining, Staining, Transgenic Assay, Mouse Assay

    Cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice were treated with 5 μM thapsigargin for 5 mins in the absence of extracellular Ca 2+ and were then exposed to 2.5 mM extracellular calcium. Upper panels show a typical time course of myocyte rounding following addition of 2.5 mM Ca 2+ in WT and TRPC3 cardiomyocytes. Data are mean ± SEM from 15 cells of at least 3 individual experiments. # = p
    Figure Legend Snippet: Cardiomyocytes from wild-type (WT) and TRPC3 transgenic mice were treated with 5 μM thapsigargin for 5 mins in the absence of extracellular Ca 2+ and were then exposed to 2.5 mM extracellular calcium. Upper panels show a typical time course of myocyte rounding following addition of 2.5 mM Ca 2+ in WT and TRPC3 cardiomyocytes. Data are mean ± SEM from 15 cells of at least 3 individual experiments. # = p

    Techniques Used: Transgenic Assay, Mouse Assay

    A) TRPC3 and B) TRPC1 protein expression in the hearts from wild-type (WT) and TRPC3 transgenic mice. Upper panels are representative immunoblots and the lower panels are mean densitometric data from 6 individual experiments normalized to β-actin. # = p
    Figure Legend Snippet: A) TRPC3 and B) TRPC1 protein expression in the hearts from wild-type (WT) and TRPC3 transgenic mice. Upper panels are representative immunoblots and the lower panels are mean densitometric data from 6 individual experiments normalized to β-actin. # = p

    Techniques Used: Expressing, Transgenic Assay, Mouse Assay, Western Blot

    4) Product Images from "The histone variant MacroH2A regulates Ca2+ influx through TRPC3 and TRPC6 channels"

    Article Title: The histone variant MacroH2A regulates Ca2+ influx through TRPC3 and TRPC6 channels

    Journal: Oncogenesis

    doi: 10.1038/oncsis.2013.40

    MacroH2A1 depletion enhances transcriptional potential of Ca 2+ binding protein-related genes. ( a ) MacroH2A1-regulated genes were analyzed by DAVID bioinformatics resources ( http://david.abcc.ncifcrf.gov ), and ontological classification of genes based on molecular function is presented as upregulated or downregulated gene groups. ( b ) For validation of microarray data, 12 genes that are related to Ca 2+ binding proteins and are upregulated in macroH2A1-depleted cells were subjected to qRT–PCR. Gapdh was used as an internal control gene. All expression values were normalized to the average of β-actin . ( c ) Trpc gene expression in control and macroH2A1-depleted LD611 cells was analyzed by qRT–PCR. ND, not detected. ( d ) Cell extracts from control and macroH2A1-depleted cells were immunoblotted with antibodies against TRPC3 and TRPC6. β-Actin was used as the internal control for loading. The analysis was performed in duplicates with comparable results. ( e ) Changes in intracellular cytosolic Ca 2+ concentration after macroH2A1 depletion were measured with the Ca 2+ -sensitive dye Fluo-8NW. ( f , g ) Control and macroH2A1-delepeted LD611 cells loaded with Fura-2 AM were stimulated with 100 μ M ATP. Representative traces of Ca 2+ in response to ATP are shown in ( f ), and changes in intracellular Ca 2+ were quantified in ( g ). ( h ) LD611 cells were stably transfected with control or macroH2A1.2 expression vectors, and the expression of macroH2A1.2 at the mRNA and protein levels was analyzed by qRT–PCR (left) and western blotting (right). ( i ) qRT–PCR was performed to check relative expressions of Ca 2+ binding-related genes, which are downregulated after macroH2A1.2 expression. ( j ) TRPC3 and TRPC6 protein levels in control and macroH2A1.2-transfected cell were evaluated by western blotting. ( k ) The intracellular Ca 2+ concentration was determined as in ( e ), but after macroH2A1.2 expression. Each bar in ( b , c , e , g – i , k ) represents the mean s.d. of three replicates in two independent experiments. * P
    Figure Legend Snippet: MacroH2A1 depletion enhances transcriptional potential of Ca 2+ binding protein-related genes. ( a ) MacroH2A1-regulated genes were analyzed by DAVID bioinformatics resources ( http://david.abcc.ncifcrf.gov ), and ontological classification of genes based on molecular function is presented as upregulated or downregulated gene groups. ( b ) For validation of microarray data, 12 genes that are related to Ca 2+ binding proteins and are upregulated in macroH2A1-depleted cells were subjected to qRT–PCR. Gapdh was used as an internal control gene. All expression values were normalized to the average of β-actin . ( c ) Trpc gene expression in control and macroH2A1-depleted LD611 cells was analyzed by qRT–PCR. ND, not detected. ( d ) Cell extracts from control and macroH2A1-depleted cells were immunoblotted with antibodies against TRPC3 and TRPC6. β-Actin was used as the internal control for loading. The analysis was performed in duplicates with comparable results. ( e ) Changes in intracellular cytosolic Ca 2+ concentration after macroH2A1 depletion were measured with the Ca 2+ -sensitive dye Fluo-8NW. ( f , g ) Control and macroH2A1-delepeted LD611 cells loaded with Fura-2 AM were stimulated with 100 μ M ATP. Representative traces of Ca 2+ in response to ATP are shown in ( f ), and changes in intracellular Ca 2+ were quantified in ( g ). ( h ) LD611 cells were stably transfected with control or macroH2A1.2 expression vectors, and the expression of macroH2A1.2 at the mRNA and protein levels was analyzed by qRT–PCR (left) and western blotting (right). ( i ) qRT–PCR was performed to check relative expressions of Ca 2+ binding-related genes, which are downregulated after macroH2A1.2 expression. ( j ) TRPC3 and TRPC6 protein levels in control and macroH2A1.2-transfected cell were evaluated by western blotting. ( k ) The intracellular Ca 2+ concentration was determined as in ( e ), but after macroH2A1.2 expression. Each bar in ( b , c , e , g – i , k ) represents the mean s.d. of three replicates in two independent experiments. * P

    Techniques Used: Binding Assay, Microarray, Quantitative RT-PCR, Expressing, Concentration Assay, Stable Transfection, Transfection, Western Blot

    Immunohistochemical staining of macroH2A1, TRPC3 and TRPC6 in tissue microarray. ( a ) Tissue microarrays containing 36 cases of bladder tumor with 12 normal tissues were subjected to immunohistochemistry with antibodies against macroH2A1, TRPC3 and TRPC6. High-power magnifications are shown for representative immunostaining samples. Bar, 50 μm. ( b ) Immunostaining scores of macroH2A1, TRPC3 and TRPC6 in bladder normal and malignant tissues. The graph indicates the percentage of sections with different scores (negative, weak, moderate and strong).
    Figure Legend Snippet: Immunohistochemical staining of macroH2A1, TRPC3 and TRPC6 in tissue microarray. ( a ) Tissue microarrays containing 36 cases of bladder tumor with 12 normal tissues were subjected to immunohistochemistry with antibodies against macroH2A1, TRPC3 and TRPC6. High-power magnifications are shown for representative immunostaining samples. Bar, 50 μm. ( b ) Immunostaining scores of macroH2A1, TRPC3 and TRPC6 in bladder normal and malignant tissues. The graph indicates the percentage of sections with different scores (negative, weak, moderate and strong).

    Techniques Used: Immunohistochemistry, Staining, Microarray, Immunostaining

    TRPC3/TRPC6 silencing results in loss of macroH2A1 function. ( a ) Cell proliferation assays were carried out in quadruplicate using cells depleted of TRPC3, TRPC6 and/or macroH2A1 as indicated. Each bar represents the mean s.d. of four replicates in three independent experiments. ( b ) Cell invasion assays were performed using cells depleted of TRPC3, TRPC6 and/or macroH2A1. Each bar represents the mean s.d. of three replicates in two independent experiments. * P
    Figure Legend Snippet: TRPC3/TRPC6 silencing results in loss of macroH2A1 function. ( a ) Cell proliferation assays were carried out in quadruplicate using cells depleted of TRPC3, TRPC6 and/or macroH2A1 as indicated. Each bar represents the mean s.d. of four replicates in three independent experiments. ( b ) Cell invasion assays were performed using cells depleted of TRPC3, TRPC6 and/or macroH2A1. Each bar represents the mean s.d. of three replicates in two independent experiments. * P

    Techniques Used:

    5) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    6) Product Images from "TRPC3 cation channel plays an important role in proliferation and differentiation of skeletal muscle myoblasts"

    Article Title: TRPC3 cation channel plays an important role in proliferation and differentiation of skeletal muscle myoblasts

    Journal: Experimental & Molecular Medicine

    doi: 10.3858/emm.2010.42.9.061

    An increase in the SR Ca 2+ content in MDG myoblasts in the presence of Pyr3, a specific blocker of TRPC3. The SR Ca 2+ of MDG myoblasts loaded with fura-2 was depleted by treatment with 10 µM CPA in the absence of extracellular Ca 2+ and presence
    Figure Legend Snippet: An increase in the SR Ca 2+ content in MDG myoblasts in the presence of Pyr3, a specific blocker of TRPC3. The SR Ca 2+ of MDG myoblasts loaded with fura-2 was depleted by treatment with 10 µM CPA in the absence of extracellular Ca 2+ and presence

    Techniques Used:

    Morphological changes in MDG/TRPC3 KD myoblasts. (A) Images of MDG/TRPC3 KD myoblasts plated on 10-cm dishes coated with collagen were obtained. Myoblasts in numbered boxes were enlarged and are presented in the lower panel (two boxes for wild-type myoblasts
    Figure Legend Snippet: Morphological changes in MDG/TRPC3 KD myoblasts. (A) Images of MDG/TRPC3 KD myoblasts plated on 10-cm dishes coated with collagen were obtained. Myoblasts in numbered boxes were enlarged and are presented in the lower panel (two boxes for wild-type myoblasts

    Techniques Used:

    Apoptotic cell death of MDG/TRPC3 KD myoblasts during exposure to differentiation conditions. (A) Fully and successfully differentiated wild-type myoblasts are shown in the upper panel. D0 to D6 means differentiation day zero to six. MDG/TRPC3 KD myoblasts
    Figure Legend Snippet: Apoptotic cell death of MDG/TRPC3 KD myoblasts during exposure to differentiation conditions. (A) Fully and successfully differentiated wild-type myoblasts are shown in the upper panel. D0 to D6 means differentiation day zero to six. MDG/TRPC3 KD myoblasts

    Techniques Used:

    TRPC3 and DHPR could be a system of checks and double-checks for skeletal myoblast differentiation by playing a redundant role
    Figure Legend Snippet: TRPC3 and DHPR could be a system of checks and double-checks for skeletal myoblast differentiation by playing a redundant role

    Techniques Used:

    Increases in both the SR Ca 2+ content and resting cytoplasmic Ca 2+ level in MDG/TRPC3 KD myoblasts. (A) the SR Ca 2+ of wild-type or MDG/TRPC3 KD myoblasts loaded with fura-2 was depleted by treatment with 10 µM cyclopiazonic acid (CPA) in the
    Figure Legend Snippet: Increases in both the SR Ca 2+ content and resting cytoplasmic Ca 2+ level in MDG/TRPC3 KD myoblasts. (A) the SR Ca 2+ of wild-type or MDG/TRPC3 KD myoblasts loaded with fura-2 was depleted by treatment with 10 µM cyclopiazonic acid (CPA) in the

    Techniques Used:

    Decreases in the expression level of TRPC4, Orai1, and MG29 in MDG/TRPC3 KD myoblasts. Solubilized cell lysate from wild-type or MDG/TRPC3 KD myoblasts (30 µg of total protein) was subjected to SDS-PAGE (10% or 12% gel) and immunoblot assay with
    Figure Legend Snippet: Decreases in the expression level of TRPC4, Orai1, and MG29 in MDG/TRPC3 KD myoblasts. Solubilized cell lysate from wild-type or MDG/TRPC3 KD myoblasts (30 µg of total protein) was subjected to SDS-PAGE (10% or 12% gel) and immunoblot assay with

    Techniques Used: Expressing, SDS Page

    Creation of the MDG/TRPC3 KD myoblast line. (A) To obtain the virus to interfere with TRPC3 mRNA in α1 S DHPR-null MDG myoblasts, a retroviral vector and two different sequences (Sequences I and II) complementary to regions of TRPC3 mRNA were used.
    Figure Legend Snippet: Creation of the MDG/TRPC3 KD myoblast line. (A) To obtain the virus to interfere with TRPC3 mRNA in α1 S DHPR-null MDG myoblasts, a retroviral vector and two different sequences (Sequences I and II) complementary to regions of TRPC3 mRNA were used.

    Techniques Used: Plasmid Preparation

    7) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    8) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    9) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    10) Product Images from "Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure"

    Article Title: Resistance to Store Depletion-induced Endothelial Injury in Rat Lung after Chronic Heart Failure

    Journal:

    doi: 10.1164/rccm.200506-847OC

    Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery
    Figure Legend Snippet: Immunostaining for transient receptor potential proteins TRPC1, TRPC3, TRPC4, and TRPC6/7 in rat lungs. Expression of each TRPC isoform in endothelium was confirmed in small muscular or partially muscularized extraalveolar vessels ( A ) and conduit artery

    Techniques Used: Immunostaining, Expressing

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Alomone Labs rabbit anti p2x3r
    Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in <t>P2X3R-containing</t> dorsal root ganglia (DRG) neurons. (A) In saline control rats,
    Rabbit Anti P2x3r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x3r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x3r - by Bioz Stars, 2022-08
    93/100 stars
      Buy from Supplier

    93
    Alomone Labs rabbit anti c terminal herg antibody
    Rescue of trafficking deficient LQT2 mutants. (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the <t>HERG</t> 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with <t>anti-C</t> terminal HERG antibody and were repeated twice (n = 2).
    Rabbit Anti C Terminal Herg Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti c terminal herg antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti c terminal herg antibody - by Bioz Stars, 2022-08
    93/100 stars
      Buy from Supplier

    95
    Alomone Labs anti trpc3
    Schematic model of <t>TRPC3</t> transmembrane domains and protein binding sites. Predicted transmembrane domains, calcium entry pore, IP 3 R, and PLCγ SH2 binding sites and deleted and substituted sites in TRPC3-DEL and TRPC3-SUB are shown. aa , amino
    Anti Trpc3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti trpc3/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti trpc3 - by Bioz Stars, 2022-08
    95/100 stars
      Buy from Supplier

    Image Search Results


    Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats,

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats,

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques:

    Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques: Expressing

    Rescue of trafficking deficient LQT2 mutants. (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: Rescue of trafficking deficient LQT2 mutants. (A) Representative western blot analysis of cell lysate from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown at 37°C and 27°C. (B) Representative western blot analysis of cell lysates from stable HEK293 cells expressing WT and traffic deficient LQT2 mutations in the HERG 1a subunit grown in the presence (+) or absence (−) of 10 µM E4031 for 36 hours. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Western Blot, Expressing

    Trafficking effects of LQT2 mutations in hetero-tetrameric HERG channels. (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: Trafficking effects of LQT2 mutations in hetero-tetrameric HERG channels. (A) Western blot analysis of cell lysates from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone, HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Mature forms of HERG 1a cmyc and HERG 1b are indicated by 155 kDa and 95 kDa labels, respectively. Immature forms of HERG 1a cmyc and HERG 1b are indicated by 135 kDa and 85 kDa labels, respectively. Equal amounts of cell lysate were loaded in all lanes as assessed using BCA protein assay quantification Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2). (B) Western blot analysis of immunoprecipitations using anti-cmyc antibody from transiently transfected HEK293 cells co-expressing WT HERG 1a cmyc tagged subunit alone, WT HERG 1b untagged alone , HERG 1a cmyc with HERG 1b or LQT2 mutant HERG 1a cmyc with HERG 1b. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Western Blot, Transfection, Expressing, Mutagenesis, BIA-KA

    Trafficking phenotype of LQT2 mutant in channels formed by HERG 1a subunit. (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Journal: PLoS ONE

    Article Title: Changes in Channel Trafficking and Protein Stability Caused by LQT2 Mutations in the PAS Domain of the HERG Channel

    doi: 10.1371/journal.pone.0032654

    Figure Lengend Snippet: Trafficking phenotype of LQT2 mutant in channels formed by HERG 1a subunit. (A) Western blot of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit. Mature fully-glycosylated HERG 1a subunit (indicated by 155 kDa label) and immature HERG 1a subunit (indicated by 135 kDa label) are indicated. (B) Western blots of membrane protein extracts from stable HEK293 cell lines expressing full-length WT or LQT2 mutant forms of HERG 1a channel subunit before (−) and after treatment (+) with N-glycosidase F. Western blots were probed with anti-C terminal HERG antibody and were repeated twice (n = 2).

    Article Snippet: Proteins were transferred by semi-dry blotting to a nitrocellulose membrane and incubated overnight at 4°C with a rabbit anti-C terminal HERG antibody (Alomone Labs) and detected by ECL.

    Techniques: Mutagenesis, Western Blot, Expressing

    Schematic model of TRPC3 transmembrane domains and protein binding sites. Predicted transmembrane domains, calcium entry pore, IP 3 R, and PLCγ SH2 binding sites and deleted and substituted sites in TRPC3-DEL and TRPC3-SUB are shown. aa , amino

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Schematic model of TRPC3 transmembrane domains and protein binding sites. Predicted transmembrane domains, calcium entry pore, IP 3 R, and PLCγ SH2 binding sites and deleted and substituted sites in TRPC3-DEL and TRPC3-SUB are shown. aa , amino

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Protein Binding, Binding Assay

    Association of TRPC3 IP 3 R binding mutants with IP 3 R. IP 3 R type II and V5-TRPC3, V5-TRPC3-DEL, or V5-TRPC3-SUB were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-V5 or anti-IP 3 R antibodies or normal rabbit

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Association of TRPC3 IP 3 R binding mutants with IP 3 R. IP 3 R type II and V5-TRPC3, V5-TRPC3-DEL, or V5-TRPC3-SUB were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-V5 or anti-IP 3 R antibodies or normal rabbit

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Binding Assay, Immunoprecipitation

    Dose response and time course of [Ca 2+ ] i after Epo stimulation of HEK 293T cells transfected with TRPC3 and Epo-R. A , Epo dose response. HEK 293T cells transfected with TRPC3 and Epo-R were stimulated with 0–40 units/ml Epo. [Ca 2+ ] i was measured

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Dose response and time course of [Ca 2+ ] i after Epo stimulation of HEK 293T cells transfected with TRPC3 and Epo-R. A , Epo dose response. HEK 293T cells transfected with TRPC3 and Epo-R were stimulated with 0–40 units/ml Epo. [Ca 2+ ] i was measured

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Transfection

    Requirement for external calcium in the Epo-stimulated calcium increase in HEK 293T cells. Fura Red-loaded HEK 293T cells were transfected with BFP-TRPC3 and Epo-R. A , cells were treated with 40 units/ml Epo in the presence (0.68 m m ) or absence (2

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Requirement for external calcium in the Epo-stimulated calcium increase in HEK 293T cells. Fura Red-loaded HEK 293T cells were transfected with BFP-TRPC3 and Epo-R. A , cells were treated with 40 units/ml Epo in the presence (0.68 m m ) or absence (2

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Transfection

    Interaction of TRPC3 with PLCγ SH2 binding site substitutions with PLCγ. PLCγ and V5-TRPC3, V5-TRPC3-F4, or V5-TRPC3-Y226F were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-PLCγ

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Interaction of TRPC3 with PLCγ SH2 binding site substitutions with PLCγ. PLCγ and V5-TRPC3, V5-TRPC3-F4, or V5-TRPC3-Y226F were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-PLCγ

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Binding Assay, Immunoprecipitation

    Western blot of HEK 293T cells transfected with siRNA targeted to PLC γ. Lysates were prepared from HEK 293T cells transfected ( Tx ' d ) with or without BFP-TRPC3 and Epo-R, and siRNA was targeted to PLCγ or control siRNA. Blots were probed

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Western blot of HEK 293T cells transfected with siRNA targeted to PLC γ. Lysates were prepared from HEK 293T cells transfected ( Tx ' d ) with or without BFP-TRPC3 and Epo-R, and siRNA was targeted to PLCγ or control siRNA. Blots were probed

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Western Blot, Transfection, Planar Chromatography

    Endogenous expression of TRPC3 in human hematopoietic cells. Western blotting was performed on lysates from UT-7 and TF-1 Epo-responsive cell lines, from CD34 + cells and from day 10 and 14 BFU-E-derived erythroblasts. Equivalent amounts of protein

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Endogenous expression of TRPC3 in human hematopoietic cells. Western blotting was performed on lysates from UT-7 and TF-1 Epo-responsive cell lines, from CD34 + cells and from day 10 and 14 BFU-E-derived erythroblasts. Equivalent amounts of protein

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Expressing, Western Blot, Derivative Assay

    Plasma membrane externalization of TRPC3 detected by cell surface biotinylation. Cell surface biotinylation was performed with HEK 293T cells expressing V5-TRPC3, V5-TRPC3-DEL, V5-TRPC3-SUB, or V5-TRPC3-F4 and Epo-R. Lysates were prepared, and immunoprecipitation

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Plasma membrane externalization of TRPC3 detected by cell surface biotinylation. Cell surface biotinylation was performed with HEK 293T cells expressing V5-TRPC3, V5-TRPC3-DEL, V5-TRPC3-SUB, or V5-TRPC3-F4 and Epo-R. Lysates were prepared, and immunoprecipitation

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Expressing, Immunoprecipitation

    Association of TRPC3 and TRPC3-F4 with PLCγ or Epo-R. A , PLCγ and V5-TRPC3 or V5-TRPC3-F4 were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-PLCγ or anti-V5 antibodies or normal rabbit

    Journal: The Journal of Biological Chemistry

    Article Title: TRPC3 Is the Erythropoietin-regulated Calcium Channel in Human Erythroid Cells *

    doi: 10.1074/jbc.M710231200

    Figure Lengend Snippet: Association of TRPC3 and TRPC3-F4 with PLCγ or Epo-R. A , PLCγ and V5-TRPC3 or V5-TRPC3-F4 were expressed in HEK 293T cells. Immunoprecipitation ( IP ) was performed on lysates with anti-PLCγ or anti-V5 antibodies or normal rabbit

    Article Snippet: Sample buffer (3×) was added to the pellets, and the samples were heated at 60 °C for 30 min. Western blotting was performed as described above, and blots were probed with anti-V5-HRP or anti-Epo-R, anti-PLCγ1, anti-IP3 R type II, anti-TRPC3, or anti-actin antibodies, followed by the appropriate HRP-conjugated secondary antibodies and ECL.

    Techniques: Immunoprecipitation

    A reduction in Ca 2+ transients in response to membrane depolarization, and the disruption of the binding between endogenous MG29 and TRPC3 in mouse primary skeletal myotubes expressing Δ116-MG29

    Journal: Biochemical and biophysical research communications

    Article Title: Interaction between mitsugumin 29 and TRPC3 participates in regulating Ca2+ transients in skeletal muscle

    doi: 10.1016/j.bbrc.2015.06.096

    Figure Lengend Snippet: A reduction in Ca 2+ transients in response to membrane depolarization, and the disruption of the binding between endogenous MG29 and TRPC3 in mouse primary skeletal myotubes expressing Δ116-MG29

    Article Snippet: For immunoblot assays, various antibodies were used: anti-RyR1, anti-DHPR, anti-SERCA1a, anti-MG29, anti-JP1, anti-JP2, and anti-GST antibodies (1:1,000) from Thermo Scientific Inc. (Rockford, IL, USA), anti-TRPC3 and anti-TRPC4 antibodies (1:800) from Alomone Laboratories (Jerusalem 9104201, Israel), and anti-Orai1, anti-STIM1, and anti-α-actin antibodies (1:1,000) from Abcam (Cambridge, MA, USA).

    Techniques: Binding Assay, Expressing

    Co-immunoprecipitation of TRPC3 with each MG29 portion

    Journal: Biochemical and biophysical research communications

    Article Title: Interaction between mitsugumin 29 and TRPC3 participates in regulating Ca2+ transients in skeletal muscle

    doi: 10.1016/j.bbrc.2015.06.096

    Figure Lengend Snippet: Co-immunoprecipitation of TRPC3 with each MG29 portion

    Article Snippet: For immunoblot assays, various antibodies were used: anti-RyR1, anti-DHPR, anti-SERCA1a, anti-MG29, anti-JP1, anti-JP2, and anti-GST antibodies (1:1,000) from Thermo Scientific Inc. (Rockford, IL, USA), anti-TRPC3 and anti-TRPC4 antibodies (1:800) from Alomone Laboratories (Jerusalem 9104201, Israel), and anti-Orai1, anti-STIM1, and anti-α-actin antibodies (1:1,000) from Abcam (Cambridge, MA, USA).

    Techniques: Immunoprecipitation