trpc6  (Alomone Labs)


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    Structured Review

    Alomone Labs trpc6
    Correlation of Ki67 expression and BrdUrd incorporation with <t>TRPC6</t> expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Trpc6, 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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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trpc6 - by Bioz Stars, 2022-07
    94/100 stars

    Images

    1) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    2) Product Images from "Store-operated Ca2+ entry in first trimester and term human placenta"

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2003.044149

    RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).
    Figure Legend Snippet: RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.
    Figure Legend Snippet: Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Techniques Used: Immunocytochemistry

    Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.
    Figure Legend Snippet: Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Techniques Used: Western Blot, Positive Control

    3) Product Images from "Store-operated Ca2+ entry in first trimester and term human placenta"

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2003.044149

    RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).
    Figure Legend Snippet: RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.
    Figure Legend Snippet: Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Techniques Used: Immunocytochemistry

    Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.
    Figure Legend Snippet: Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Techniques Used: Western Blot, Positive Control

    4) Product Images from "Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts"

    Article Title: Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.116.003911

    Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.
    Figure Legend Snippet: Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.

    Techniques Used: Activity Assay, Expressing, Staining, Immunoprecipitation

    5) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    6) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    7) Product Images from "Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle"

    Article Title: Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00629.2008

    Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5
    Figure Legend Snippet: Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5

    Techniques Used:

    A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1
    Figure Legend Snippet: A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1

    Techniques Used: Expressing

    A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time
    Figure Legend Snippet: A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time

    Techniques Used: Expressing

    A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as
    Figure Legend Snippet: A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as

    Techniques Used: Expressing, Small Interfering RNA

    8) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    9) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    10) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    11) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    12) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    13) Product Images from "Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle"

    Article Title: Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00629.2008

    Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5
    Figure Legend Snippet: Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5

    Techniques Used:

    A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1
    Figure Legend Snippet: A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1

    Techniques Used: Expressing

    A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time
    Figure Legend Snippet: A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time

    Techniques Used: Expressing

    A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as
    Figure Legend Snippet: A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as

    Techniques Used: Expressing, Small Interfering RNA

    14) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    15) Product Images from "Store-operated Ca2+ entry in first trimester and term human placenta"

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2003.044149

    RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).
    Figure Legend Snippet: RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.
    Figure Legend Snippet: Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Techniques Used: Immunocytochemistry

    Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.
    Figure Legend Snippet: Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Techniques Used: Western Blot, Positive Control

    16) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    17) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    18) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    19) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    20) Product Images from "Cystic fibrosis transmembrane conductance regulator dysfunction in platelets drives lung hyperinflammation"

    Article Title: Cystic fibrosis transmembrane conductance regulator dysfunction in platelets drives lung hyperinflammation

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI129635

    Calcium entry measured by the ratiometric Indo-1 assay in thrombin-stimulated platelets from mice and humans. ( A ) Kinetic tracings of Indo-1 Violet/Blue MFI measured in platelets isolated from CFTR –/– (blue), TRPC6 –/– (green), and WT (red) mice with 0.125 IU thrombin introduced at 60-second intervals starting at 30 seconds (black arrows). ( B ) Peak MFI in WT, CFTR –/– , and TRPC6 –/– platelets incubated with vehicles, CF172, or CF172 plus SK. Data are mean ± SEM of 7 to 11 animals per group. ( C ) Peak MFI measured in platelets isolated from healthy human and CF subjects not on modulators incubated with vehicles, CF172, or CF172 plus SK. ( D ) CD62P and ( E ) PAC-1 expression in platelets from human controls, CF platelets plus modulators (lumacaftor/ivacaftor), and CF platelets not treated with modulators (no modulators). Data are presented as minimum-to-maximum whiskers and box plots showing the median and interquartile ranges. ( C – E ) n = 6–12 subjects per group. Data in B – E were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.
    Figure Legend Snippet: Calcium entry measured by the ratiometric Indo-1 assay in thrombin-stimulated platelets from mice and humans. ( A ) Kinetic tracings of Indo-1 Violet/Blue MFI measured in platelets isolated from CFTR –/– (blue), TRPC6 –/– (green), and WT (red) mice with 0.125 IU thrombin introduced at 60-second intervals starting at 30 seconds (black arrows). ( B ) Peak MFI in WT, CFTR –/– , and TRPC6 –/– platelets incubated with vehicles, CF172, or CF172 plus SK. Data are mean ± SEM of 7 to 11 animals per group. ( C ) Peak MFI measured in platelets isolated from healthy human and CF subjects not on modulators incubated with vehicles, CF172, or CF172 plus SK. ( D ) CD62P and ( E ) PAC-1 expression in platelets from human controls, CF platelets plus modulators (lumacaftor/ivacaftor), and CF platelets not treated with modulators (no modulators). Data are presented as minimum-to-maximum whiskers and box plots showing the median and interquartile ranges. ( C – E ) n = 6–12 subjects per group. Data in B – E were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

    Techniques Used: Mouse Assay, Isolation, Incubation, Expressing

    Characterization of TRPC6 in platelets. ( A , B ) mRNA expression of TRPC isoforms TRPC1 and TRPC6 in platelets from WT, CFTR –/– , CF fl/fl , CF-LysM, and CF-PF4 mice. TRPC isoforms 2, 3, 4, 5, and 7 were undetectable (not shown). ( C ) Immunofluorescence staining and ( D ) flow cytometry analysis of CD41 (red) and TRPC6 (blue) in platelets from WT and TRPC6 –/– mice (representative of 3 independent experiments). Scale bar: 2.5 μm. ( E and F ) CD62P expression on platelets from ( E ) WT, CFTR –/– , TRPC6 –/– , and CFTR –/– × TRPC6 –/– mice, and ( F ) CF fl/fl and CF-PF4 mice after thrombin challenge with or without incubation with vehicles, CF172, or CF172 plus SKF-96365 (SK). Data are mean ± SEM of 5 to 11 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; **** P ≤ 0.0001.
    Figure Legend Snippet: Characterization of TRPC6 in platelets. ( A , B ) mRNA expression of TRPC isoforms TRPC1 and TRPC6 in platelets from WT, CFTR –/– , CF fl/fl , CF-LysM, and CF-PF4 mice. TRPC isoforms 2, 3, 4, 5, and 7 were undetectable (not shown). ( C ) Immunofluorescence staining and ( D ) flow cytometry analysis of CD41 (red) and TRPC6 (blue) in platelets from WT and TRPC6 –/– mice (representative of 3 independent experiments). Scale bar: 2.5 μm. ( E and F ) CD62P expression on platelets from ( E ) WT, CFTR –/– , TRPC6 –/– , and CFTR –/– × TRPC6 –/– mice, and ( F ) CF fl/fl and CF-PF4 mice after thrombin challenge with or without incubation with vehicles, CF172, or CF172 plus SKF-96365 (SK). Data are mean ± SEM of 5 to 11 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; **** P ≤ 0.0001.

    Techniques Used: Expressing, Mouse Assay, Immunofluorescence, Staining, Flow Cytometry, Incubation

    Lung injury measurements in CFTR and TRPC6 mutant mice after intratracheal LPS or PAO1. ( A ) BAL WBCs, ( B ) neutrophils, ( C ) total protein, ( D ) thromboxane B 2, ( E ) NETs (NE-DNA ELISA), and ( F ) NETs (citH3-DNA ELISA) in CFTR × TRPC6 mutant mice (genotypes indicated in x axis label). Data are mean ± SEM of 5 to 6 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. ( G – J ) Lung injury and bacterial counts after intratracheal PAO1. ( G ) BAL WBCs, ( H ) neutrophils, ( I ) total protein, and ( J ) lung colonies in CFTR and TRPC6 mutant mice. Data are mean ± SEM of 5 to 6 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.
    Figure Legend Snippet: Lung injury measurements in CFTR and TRPC6 mutant mice after intratracheal LPS or PAO1. ( A ) BAL WBCs, ( B ) neutrophils, ( C ) total protein, ( D ) thromboxane B 2, ( E ) NETs (NE-DNA ELISA), and ( F ) NETs (citH3-DNA ELISA) in CFTR × TRPC6 mutant mice (genotypes indicated in x axis label). Data are mean ± SEM of 5 to 6 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. ( G – J ) Lung injury and bacterial counts after intratracheal PAO1. ( G ) BAL WBCs, ( H ) neutrophils, ( I ) total protein, and ( J ) lung colonies in CFTR and TRPC6 mutant mice. Data are mean ± SEM of 5 to 6 animals per group. Data were analyzed by 2-way ANOVA. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

    Techniques Used: Mutagenesis, Mouse Assay, Enzyme-linked Immunosorbent Assay

    21) Product Images from "Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts"

    Article Title: Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.116.003911

    Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.
    Figure Legend Snippet: Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.

    Techniques Used: Activity Assay, Expressing, Staining, Immunoprecipitation

    22) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    23) Product Images from "BMP4 Increases Canonical Transient Receptor Potential Protein Expression by Activating p38 MAPK and ERK1/2 Signaling Pathways in Pulmonary Arterial Smooth Muscle Cells"

    Article Title: BMP4 Increases Canonical Transient Receptor Potential Protein Expression by Activating p38 MAPK and ERK1/2 Signaling Pathways in Pulmonary Arterial Smooth Muscle Cells

    Journal: American Journal of Respiratory Cell and Molecular Biology

    doi: 10.1165/rcmb.2012-0051OC

    SB203580 and PD98059 inhibited BMP4-induced transient receptor potential (TRPC)–1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with DMSO, p38 inhibitor SB203580 (10 μM), or ERK1/2 inhibitor PD98059 (5 μM)
    Figure Legend Snippet: SB203580 and PD98059 inhibited BMP4-induced transient receptor potential (TRPC)–1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with DMSO, p38 inhibitor SB203580 (10 μM), or ERK1/2 inhibitor PD98059 (5 μM)

    Techniques Used: Expressing

    Effects of ERK1/2 siRNA on BMP4-induced TRPC1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with ERK1/2 siRNA (50 nM) or an equal amount of nontargeting control siRNA (NT siRNA) for 24 hours, and were then incubated with BMP4
    Figure Legend Snippet: Effects of ERK1/2 siRNA on BMP4-induced TRPC1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with ERK1/2 siRNA (50 nM) or an equal amount of nontargeting control siRNA (NT siRNA) for 24 hours, and were then incubated with BMP4

    Techniques Used: Expressing, Incubation

    Effects of p38 small interfering RNA (siRNA) on BMP4-induced TRPC1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with p38 siRNA (50 nM) or an equal amount of nontargeting control siRNA (NT siRNA) for 24 hours, and were then
    Figure Legend Snippet: Effects of p38 small interfering RNA (siRNA) on BMP4-induced TRPC1, TRPC4, and TRPC6 expression in rat distal PASMCs. Cells were pretreated with p38 siRNA (50 nM) or an equal amount of nontargeting control siRNA (NT siRNA) for 24 hours, and were then

    Techniques Used: Small Interfering RNA, Expressing

    24) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    25) Product Images from "Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension"

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0405908101

    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Figure Legend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Techniques Used: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P
    Figure Legend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Techniques Used: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P
    Figure Legend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Techniques Used: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.
    Figure Legend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Techniques Used: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    26) Product Images from "Store-operated Ca2+ entry in first trimester and term human placenta"

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2003.044149

    RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).
    Figure Legend Snippet: RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.
    Figure Legend Snippet: Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Techniques Used: Immunocytochemistry

    Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.
    Figure Legend Snippet: Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Techniques Used: Western Blot, Positive Control

    27) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    28) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    29) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    30) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    31) Product Images from "Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle"

    Article Title: Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00629.2008

    Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5
    Figure Legend Snippet: Effects of TRPC1 and TRPC6 knockdown on hypoxic increases of SOCE in PASMCs as assessed by [Ca 2+ ] restoration ( A and B ) and Mn 2+ quenching ( C and D ) are shown. A : representative traces of [Ca 2+ ] i responses to restoration of extracellular [Ca 2+ ] to 2.5

    Techniques Used:

    A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1
    Figure Legend Snippet: A and B : expression of TRPC1 ( A ) and TRPC6 ( B ) mRNA relative to 18s in distal pulmonary arteries (PA) from rats exposed to normoxia or CH (10% O 2 for 21 days) and treated with or without sildenafil (50 mg · kg −1 · day −1

    Techniques Used: Expressing

    A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time
    Figure Legend Snippet: A and B : expression of canonical transient receptor potential 1 (TRPC1; A ) and 6 (TRPC6; B ) mRNA relative to 18s in PASMCs treated with sildenafil (300 nM) or vehicle (control) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as determined by real-time

    Techniques Used: Expressing

    A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as
    Figure Legend Snippet: A : expression of TRPC1 and TRPC6 mRNA relative to 18s in PASMCs treated with nontargeting control small interfering RNA (siNT), siRNA targeted to TRPC1 (siT1), or siRNA targeted to TRPC6 (siT6) for 60 h under normoxic or hypoxic (4% O 2 ) conditions as

    Techniques Used: Expressing, Small Interfering RNA

    32) Product Images from "Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts"

    Article Title: Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.116.003911

    Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.
    Figure Legend Snippet: Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.

    Techniques Used: Activity Assay, Expressing, Staining, Immunoprecipitation

    33) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    34) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    35) Product Images from "ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin"

    Article Title: ATRA attenuate proteinuria via downregulation of TRPC6 in glomerulosclerosis rats induced by adriamycin

    Journal: Renal Failure

    doi: 10.1080/0886022X.2018.1456459

    Evaluation of protein expression of TRPC6 (Western-blot). ※ p
    Figure Legend Snippet: Evaluation of protein expression of TRPC6 (Western-blot). ※ p

    Techniques Used: Expressing, Western Blot

    The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.
    Figure Legend Snippet: The typical immunohistochemical staining of TGF-β, Col-IV and TRPC6 in glomerulus in three groups. Stainings for TGF-β, Col-IV, and TRPC6 in the GS group were markedly increased when compared with those in the SHO group. The positive stainings of TGF-β, Col-IV, and TRPC6 in the ATRA group were markedly reduced when compared with the GS group. SHO: sham operation group; GS: GS model group; ATRA: GS model group treated with ATRA. Magnification 400×.

    Techniques Used: Immunohistochemistry, Staining

    36) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

    37) Product Images from "Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction"

    Article Title: Role of phosphatase and tensin homolog in hypoxic pulmonary vasoconstriction

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx076

    Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia causes ROCK-dependent translocation of TRPC6 and PTEN to caveolae. Caveolar fractions were isolated from PASMC (input: whole cell lysate) by sucrose gradient centrifugation, and identified by the presence of the marker protein caveolin-1. Representative immunoblots show ( A ) absence of PTEN and TRPC6 from caveolae of normoxic PASMC, but ( B ) recruitment to caveolar fractions within 15 min of hypoxia (1% O 2 ), that was ( C ) blocked by Y27632 (5 µMol/L). ( D ) Group data from n = 3–5 independent experiments each show caveolar recruitment of TRPC6 and PTEN, expressed as fraction of protein detected in caveolar fractions. Group data are means ± SEM, * P

    Techniques Used: Translocation Assay, Isolation, Gradient Centrifugation, Marker, Western Blot

    Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.
    Figure Legend Snippet: Schematic outline of the proposed role of PTEN in HPV: Hypoxia, either directly and/or via formation of S1P, activates ROCKin PASMC, which mediates the interaction between PTEN and TRPC6 and their translocation to caveolae, where TRPC6 facilitates Ca 2+ entry and subsequent PASMC contraction.

    Techniques Used: Translocation Assay

    Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P
    Figure Legend Snippet: Hypoxia increases the interaction of PTEN with TRPC6 in a ROCK-dependent manner. ( A ) PASMCs were exposed to either normoxia (n), hypoxia (h; 1% O 2 ), or hypoxia in the presence of Y27632 (Y; 5µMol/L), or to S1P (S; 10 µMol/L) for 5 min. Representative immunoblots show TRPC6 and PTEN expression in PASMC for whole cell lysate (input) and after immunoprecipitation for PTEN (Co-IP). Group data from n = 5–7 independent replicates showing TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel, demonstrate increased interaction of PTEN with TRPC6 in response to hypoxia and S1P, respectively. ( B ) Isolated lung were ventilated with normoxic or hypoxic (1% O 2 ) gas for 3 min, and tissue was snap-frozen and lysed. Representative immunoblots show TRPC6 and PTEN expression after immunoprecipitation for PTEN; group data from n = 6 replicates show quantification of TRPC6-over-PTEN ratio normalized to the normoxic control group from the same gel. ( C ) Representative images show PLA for the interaction between PTEN and TRPC6 in PASMC following exposure to either normoxia, hypoxia, or S1P(10 µMol/L) for 5 min, in the presence or absence of Y27632 (5 µMol/L), or LPA (3 µMol/L) for 15 min. Red puncta indicate sites of interaction between PTEN and TRPC6, nuclei are counterstained in blue with DAPI. ( D ) Group data show quantification of the PLA from eight cells from three independent experiments each. Group data are means ± SEM, * P

    Techniques Used: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Isolation, Proximity Ligation Assay

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    Alomone Labs trpc6
    Correlation of Ki67 expression and BrdUrd incorporation with <t>TRPC6</t> expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)
    Trpc6, 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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    Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    doi: 10.1073/pnas.0405908101

    Figure Lengend Snippet: Correlation of Ki67 expression and BrdUrd incorporation with TRPC6 expression. ( A ) Dot-plot analysis (representative of three trials) of TRPC6 expression vs. Ki67 expression in growth-arrested and proliferating PASMC. Upper right quadrants represent cells in the S phase. ( B ) Three-color dot-plot analysis of TRPC6 distribution vs. BrdUrd incorporation and Ki67 expression in proliferating PASMC (representative of three trials). The box in Left indicates the S-phase cells that are stained by both Ki67 and BrdUrd. ( C ) Fluorescent images (×40) showing PASMC stained by Ki67 and TRPC6. An overlay of the two fluorescent images identifies PASMC expressing both Ki67 and TRPC6. Yellow and green arrows indicate Ki67-positive and -negative PASMC, respectively. (Scale bar, 20 μm.)

    Article Snippet: Furthermore, a majority of S-phase cells (double-labeled by BrdUrd and Ki67) also expressed TRPC6 , indicating that its expression is elevated during DNA synthesis.

    Techniques: Expressing, Staining

    TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    doi: 10.1073/pnas.0405908101

    Figure Lengend Snippet: TRPC6 siRNA attenuates TRPC6 expression and proliferation in IPAH cells. ( A ) Effect of two concentrations of TRPC6-siRNA-2 expression vector, and its scrambled control (4 μg/ml), on TRPC6 protein expression. ( B ) ( Left ) Normalized [ 3 H]thymidine uptake plotted for cells untreated (Cont) or treated with the scrambled siRNA duplex (Scram, 200 nM). ( Right ) Cell growth in cells transfected with two concentrations of either the scrambled ( n = 8) or TRPC6 ( n = 8) siRNA duplexes. Data are normalized to the basal [ 3 H]thymidine uptake in the absence of siRNA (Cont). *** , P

    Article Snippet: Furthermore, a majority of S-phase cells (double-labeled by BrdUrd and Ki67) also expressed TRPC6 , indicating that its expression is elevated during DNA synthesis.

    Techniques: Expressing, Plasmid Preparation, Transfection

    TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    doi: 10.1073/pnas.0405908101

    Figure Lengend Snippet: TRPC6 mRNA and protein expression is increased in pulmonary tissues from IPAH patients. ( A ) Representative and summarized (NPH, n = 3; SPH, n = 8; IPAH, n = 3; normalized to α-actin) TRPC6 protein expression in SPH, NPH, and IPAH lung tissues and their α-actin controls. ( B ) Representative and summarized ( n = 3 for each) data showing TRPC6 mRNA products in isolated NPH-, SPH-, or IPAH-PASMC. M, 100-bp DNA ladder. ( C ) Representative and summarized ( n = 5 for each) TRPC6 protein expression in isolated PASMC from NPH, SPH, and IPAH patients. ( D ) Representative ( Top , ×40; Middle , ×100) and summarized immunofluorescence staining of FITC-conjugated TRPC6 ( Bottom ) in isolated NPH-( n = 11), SPH ( n = 21), and IPAH-( n = 36) PASMC. (Scale bars, 20 μm.) *** , P

    Article Snippet: Furthermore, a majority of S-phase cells (double-labeled by BrdUrd and Ki67) also expressed TRPC6 , indicating that its expression is elevated during DNA synthesis.

    Techniques: Expressing, Isolation, Immunofluorescence, Staining

    Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension

    doi: 10.1073/pnas.0405908101

    Figure Lengend Snippet: Cell cycle analysis of growth-arrested and proliferating PASMC and correlation of TRPC6 expression with PASMC proliferation. ( A ) Flow cytometry graphs (representative of three trials) of cell cycle analysis for growth-arrested (SMBM) and proliferating (SMGM) PASMC. Number of cells is plotted as a function of DNA content. ( B ) Graphs showing cell number as a function of Ki67 expression, BrdUrd incorporation, or TRPC6 expression in SMBM- and SMGM-treated PASMC.

    Article Snippet: Furthermore, a majority of S-phase cells (double-labeled by BrdUrd and Ki67) also expressed TRPC6 , indicating that its expression is elevated during DNA synthesis.

    Techniques: Cell Cycle Assay, Expressing, Flow Cytometry, Cytometry

    RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Journal: The Journal of Physiology

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    doi: 10.1113/jphysiol.2003.044149

    Figure Lengend Snippet: RT-PCR detection of TRPC mRNA RT-PCR was performed using gene-specific primers for A , TRPC1, B , TRPC3, C , TRPC4, D , TRPC5, E , TRPC6 and F , β-actin. PCR product sizes are indicated. L, 100 bp ladder; −, negative control (H 2 O); SI, small intestine (Clontech); HB, human brain (Clontech); I, first trimester placenta ( n = 7 pooled); II, second trimester placentas ( n = 5 pooled); III, term placentas ( n = 8 pooled).

    Article Snippet: At the protein level we examined expression of TRPC1, TRPC3, TRPC4 and TRPC6 and found that term placenta expressed TRPC3, TRPC4 and TRPC6 but not TRPC1.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control

    Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Journal: The Journal of Physiology

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    doi: 10.1113/jphysiol.2003.044149

    Figure Lengend Snippet: Immunocytochemistry showing localisation for TRPC3, TRPC4 and TRPC6 in first trimester ( A , C and E ) and term placenta ( B , D and F ) Negative controls for first trimester ( G ) and term placenta ( H ) are also shown. Samples were magnified × 1000; a scale bar is given in H . S, syncytiotrophoblast; C, cytotrophoblast; VC, villous core; M, microvillous membrane; B, basal membrane.

    Article Snippet: At the protein level we examined expression of TRPC1, TRPC3, TRPC4 and TRPC6 and found that term placenta expressed TRPC3, TRPC4 and TRPC6 but not TRPC1.

    Techniques: Immunocytochemistry

    Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Journal: The Journal of Physiology

    Article Title: Store-operated Ca2+ entry in first trimester and term human placenta

    doi: 10.1113/jphysiol.2003.044149

    Figure Lengend Snippet: Western blotting for TRPC1 ( A , B ), TRPC3 ( C , D ) and TRPC6 ( E , F ) Protein sizes are given. Preabsorption of each primary antibody with antigen (Ag) shows removal of specific bands ( A , C , E ). ECL detection was for 30 min for preabsorption studies and 1 h for comparison of first trimester and term tissue. RB, rat brain (positive control tissue); T, term placenta; F, first trimester placenta. In B , D , F , and G each lane corresponds to samples prepared from four different first trimester and term placentas.

    Article Snippet: At the protein level we examined expression of TRPC1, TRPC3, TRPC4 and TRPC6 and found that term placenta expressed TRPC3, TRPC4 and TRPC6 but not TRPC1.

    Techniques: Western Blot, Positive Control

    Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Tadalafil Treatment Delays the Onset of Cardiomyopathy in Dystrophin‐Deficient Hearts

    doi: 10.1161/JAHA.116.003911

    Figure Lengend Snippet: Tadalafil (Tad) modulates TRPC 6 and protease levels and activity in golden retriever muscular dystrophy ( GRMD) left ventricle (LV). A, Gene expression of Pde5 , Trpc3 , and Trpc6 in control and Tad‐treated GRMD LV (normalized to Gapdh and displayed as arbitrary units (AU) relative to untreated GRMD values). All values and individual values are presented, and group values are represented as mean± SD . Representative blots of Tad‐induced changes in TRPC 6, μ‐calpain, m‐calpain, cardiac troponin I ( cTnI) , spectrin 150‐kDa cleavage product, α‐actinin 80‐kDa cleavage product, integrin β1 75‐kDa cleavage product, dysferlin C72 fragment, full‐length utrophin, and fibronectin protein content (B), along with percentage difference between untreated and treated values (Cohen's d shown to display effect size). Ponceau Red staining is used as a loading control. C, TRPC 6 and TRPC 3 immunoprecipitation (IP) results for phosphorylated threonine (p‐Thr) with blot density quantified as phopho‐ to total (p/t) protein ratio.

    Article Snippet: It appears that this basal PDE5 activity is sufficient to modulate both TRPC6 and m‐calpain.

    Techniques: Activity Assay, Expressing, Staining, Immunoprecipitation