capsazepine  (Alomone Labs)


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    Alomone Labs capsazepine
    Capsazepine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capsazepine/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    capsazepine - by Bioz Stars, 2022-12
    92/100 stars

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    Alomone Labs rabbit polyclonal anti trpc6 antibody
    Melatonin downregulates <t>TRPC6</t> expression in MDA-MB-231 cells. MCF10A ( A , E and I ) and MDA-MB-231 cells ( B – D , F – G and J – L ) were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with the anti-TRPC6 ( A , B , E , F , I , and J ), anti-PMCA ( C , G , and K ), anti-Orai1 ( D , H and L ), or anti-β-actin antibodies, as described in Experimental procedures . Blots are representative of five to ten separate experiments. Scatter plots represent TRPC6, Orai1, or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (0.001
    Rabbit Polyclonal Anti Trpc6 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti trpc6 antibody/product/Alomone Labs
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti trpc6 antibody - by Bioz Stars, 2022-12
    93/100 stars
      Buy from Supplier

    92
    Alomone Labs capsazepine
    Melatonin downregulates <t>TRPC6</t> expression in MDA-MB-231 cells. MCF10A ( A , E and I ) and MDA-MB-231 cells ( B – D , F – G and J – L ) were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with the anti-TRPC6 ( A , B , E , F , I , and J ), anti-PMCA ( C , G , and K ), anti-Orai1 ( D , H and L ), or anti-β-actin antibodies, as described in Experimental procedures . Blots are representative of five to ten separate experiments. Scatter plots represent TRPC6, Orai1, or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (0.001
    Capsazepine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capsazepine/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    capsazepine - by Bioz Stars, 2022-12
    92/100 stars
      Buy from Supplier

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    Melatonin downregulates TRPC6 expression in MDA-MB-231 cells. MCF10A ( A , E and I ) and MDA-MB-231 cells ( B – D , F – G and J – L ) were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with the anti-TRPC6 ( A , B , E , F , I , and J ), anti-PMCA ( C , G , and K ), anti-Orai1 ( D , H and L ), or anti-β-actin antibodies, as described in Experimental procedures . Blots are representative of five to ten separate experiments. Scatter plots represent TRPC6, Orai1, or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (0.001

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Melatonin downregulates TRPC6 expression in MDA-MB-231 cells. MCF10A ( A , E and I ) and MDA-MB-231 cells ( B – D , F – G and J – L ) were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with the anti-TRPC6 ( A , B , E , F , I , and J ), anti-PMCA ( C , G , and K ), anti-Orai1 ( D , H and L ), or anti-β-actin antibodies, as described in Experimental procedures . Blots are representative of five to ten separate experiments. Scatter plots represent TRPC6, Orai1, or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (0.001

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Expressing, Multiple Displacement Amplification, SDS Page, Western Blot

    Overexpression of TRPC6 attenuates melatonin-evoked inhibition of store-operated Ca 2+ entry in MDA-MB-231 cells. A , left panel , MDA-MB-231 cells were transfected with TRPC6dn-YFP or TRPC6-YFP expression plasmids. Forty-eight hours later, TRPC6 expression was determined by fluorescence microscopy. A , right panel , MDA-MB-231 cells were transfected with TRPC6dn-YFP or TRPC6-YFP expression plasmids or empty vector (control). Forty-eight hours later cells were loaded with fura-2. Fura-2-loaded cells were perfused with a Ca 2+ -free medium (100 μM EGTA added) and then stimulated with TG (1 μM) followed by reintroduction of external Ca 2+ (final concentration 1 mM) to initiate Ca 2+ entry. Scatter plots represent the quantification of TG-evoked Ca 2+ release and entry determined as described in Experimental procedures . Data are presented as mean ± SD. Dots represent a single experiment including 20–30 cells. Analysis of statistical significance was performed using one-way ANOVA (F = 1.82; p = 0.19 for Ca 2+ release and F = 546.5 and p

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Overexpression of TRPC6 attenuates melatonin-evoked inhibition of store-operated Ca 2+ entry in MDA-MB-231 cells. A , left panel , MDA-MB-231 cells were transfected with TRPC6dn-YFP or TRPC6-YFP expression plasmids. Forty-eight hours later, TRPC6 expression was determined by fluorescence microscopy. A , right panel , MDA-MB-231 cells were transfected with TRPC6dn-YFP or TRPC6-YFP expression plasmids or empty vector (control). Forty-eight hours later cells were loaded with fura-2. Fura-2-loaded cells were perfused with a Ca 2+ -free medium (100 μM EGTA added) and then stimulated with TG (1 μM) followed by reintroduction of external Ca 2+ (final concentration 1 mM) to initiate Ca 2+ entry. Scatter plots represent the quantification of TG-evoked Ca 2+ release and entry determined as described in Experimental procedures . Data are presented as mean ± SD. Dots represent a single experiment including 20–30 cells. Analysis of statistical significance was performed using one-way ANOVA (F = 1.82; p = 0.19 for Ca 2+ release and F = 546.5 and p

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Over Expression, Inhibition, Multiple Displacement Amplification, Transfection, Expressing, Fluorescence, Microscopy, Plasmid Preparation, Concentration Assay

    Effect of melatonin on calcium mobilization, viability, and migration in MDA-MB-468 cells. A , MDA-MB-468 cells were treated for 72 h with melatonin (100–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with anti-TRPC6, anti-PMCA, anti-Orai1, or anti-β-actin antibody, as described in Experimental procedures . Blots are representative of five to six separate experiments. Scatter plots represent TRPC6, Orai1 or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (F = 10.72, 0.16, and 0.85 and p = 0.001, 0.84, and 0.44 for TRPC6, PMCA, and Orai1, respectively) with post-hoc Dunnett's test for TRPC6 (∗ p

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Effect of melatonin on calcium mobilization, viability, and migration in MDA-MB-468 cells. A , MDA-MB-468 cells were treated for 72 h with melatonin (100–1000 nM) or the vehicle (control) and lysed. Whole-cell lysates were subjected to 10% SDS-PAGE and western blotting with anti-TRPC6, anti-PMCA, anti-Orai1, or anti-β-actin antibody, as described in Experimental procedures . Blots are representative of five to six separate experiments. Scatter plots represent TRPC6, Orai1 or PMCA expression. Analysis of statistical significance was performed using one-way ANOVA (F = 10.72, 0.16, and 0.85 and p = 0.001, 0.84, and 0.44 for TRPC6, PMCA, and Orai1, respectively) with post-hoc Dunnett's test for TRPC6 (∗ p

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Migration, Multiple Displacement Amplification, SDS Page, Western Blot, Expressing

    Effect of pharmacological inhibition or silencing of TRPC6 on Ca 2+ entry in the presence of melatonin in MDA-MB-231 cells. A , Fura-2-loaded MDA-MB-231 cells were pretreated for 10 min with SAR7334 (1 μM). Cells were then stimulated in the presence of 1 mM extracellular Ca 2+ with OAG (100 μM). Scatter plots represent the quantification of OAG-induced Ca 2+ entry determined as described in Experimental procedures . Data are presented as mean ± SD. Dots represent a single experiment including 20–30 cells. Analysis of statistical significance was performed using Student's t -test (∗ p

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Effect of pharmacological inhibition or silencing of TRPC6 on Ca 2+ entry in the presence of melatonin in MDA-MB-231 cells. A , Fura-2-loaded MDA-MB-231 cells were pretreated for 10 min with SAR7334 (1 μM). Cells were then stimulated in the presence of 1 mM extracellular Ca 2+ with OAG (100 μM). Scatter plots represent the quantification of OAG-induced Ca 2+ entry determined as described in Experimental procedures . Data are presented as mean ± SD. Dots represent a single experiment including 20–30 cells. Analysis of statistical significance was performed using Student's t -test (∗ p

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Inhibition, Multiple Displacement Amplification

    Effect of melatonin on TRPC6 synthesis and degradation in MDA-MB-231 cells. A and B , MDA-MB-231 cells were treated for 48 or 72 h with melatonin (100 nM) or the vehicle (control). The expression of TRPC6 ( A ) or miR26a-5p ( B ) was determined as described in Experimental procedures . Scatter plots represent relative expression as compared with control (n = 4–7). Analysis of statistical significance was performed using one-way ANOVA (F and p values were 0.41 and 0.74, respectively, for [ A ] and 0.17 and 0.91, respectively, for [ B ]). C , MDA-MB-231 cells were treated for 72 h with melatonin (100 nM) or the vehicle (control) and lysed. Sixteen hours before the end of the treatment, cells were treated with 10 μM MG132 or the vehicle. Whole-cell lysates were immunoprecipitated with anti-TRPC6 antibody. Immunoprecipitates were subjected to 10% SDS-PAGE and subsequent western blotting with specific anti-TRPC6 (left panel), antiubiquitin (right panel), or anti-β-actin antibody, as indicated. The panels show results from one experiment representative of three others. Molecular masses indicated on the right were determined using molecular-mass markers run in the same gel. Bar graphs represent the expression of TRPC6 (left panel) and the quantification of TRPC6-associated ubiquitin. Results are presented as arbitrary optical density units and expressed as percentage of control. Analysis of statistical significance was performed using one-way ANOVA (F = 14.71 ( p = 0.0003) and 71.28 ( p

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Effect of melatonin on TRPC6 synthesis and degradation in MDA-MB-231 cells. A and B , MDA-MB-231 cells were treated for 48 or 72 h with melatonin (100 nM) or the vehicle (control). The expression of TRPC6 ( A ) or miR26a-5p ( B ) was determined as described in Experimental procedures . Scatter plots represent relative expression as compared with control (n = 4–7). Analysis of statistical significance was performed using one-way ANOVA (F and p values were 0.41 and 0.74, respectively, for [ A ] and 0.17 and 0.91, respectively, for [ B ]). C , MDA-MB-231 cells were treated for 72 h with melatonin (100 nM) or the vehicle (control) and lysed. Sixteen hours before the end of the treatment, cells were treated with 10 μM MG132 or the vehicle. Whole-cell lysates were immunoprecipitated with anti-TRPC6 antibody. Immunoprecipitates were subjected to 10% SDS-PAGE and subsequent western blotting with specific anti-TRPC6 (left panel), antiubiquitin (right panel), or anti-β-actin antibody, as indicated. The panels show results from one experiment representative of three others. Molecular masses indicated on the right were determined using molecular-mass markers run in the same gel. Bar graphs represent the expression of TRPC6 (left panel) and the quantification of TRPC6-associated ubiquitin. Results are presented as arbitrary optical density units and expressed as percentage of control. Analysis of statistical significance was performed using one-way ANOVA (F = 14.71 ( p = 0.0003) and 71.28 ( p

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Multiple Displacement Amplification, Expressing, Immunoprecipitation, SDS Page, Western Blot

    Expression of exogenous TRPC6 reverses the antiproliferative effect of melatonin in MDA-MB-231 cells. MDA-MB-231 cells were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control). Forty-eight hours before the end of the treatment period, cells were transfected with TRPC6-YFP ( B , E , and H ) or TRPC6dn-YFP ( C , F , and I ) expression plasmid or empty vector ( A , D and G ). Cell proliferation was assessed at time = 0, 24, 48, and 72 h using the BrdU cell proliferation assay kit, as described in Experimental procedures . Scatter plots represent cell proliferation 0, 24, 48, and 72 h after the onset of the experiment, presented as BrdU uptake rate (n = 6). Analysis of statistical significance was performed using two-way ANOVA (for A , D , G : 0 h, F values were 0.14, 0.14, and 1.35 and p values were 0.93, 0.86, and 0.24 for concentration, time, and the interaction respectively; for A , D , G : 24 h, F values were 593.8, 1570, and 199.6 and p values were

    Journal: The Journal of Biological Chemistry

    Article Title: Melatonin downregulates TRPC6, impairing store-operated calcium entry in triple-negative breast cancer cells

    doi: 10.1074/jbc.RA120.015769

    Figure Lengend Snippet: Expression of exogenous TRPC6 reverses the antiproliferative effect of melatonin in MDA-MB-231 cells. MDA-MB-231 cells were treated for 48, 72, and 168 h with melatonin (10–1000 nM) or the vehicle (control). Forty-eight hours before the end of the treatment period, cells were transfected with TRPC6-YFP ( B , E , and H ) or TRPC6dn-YFP ( C , F , and I ) expression plasmid or empty vector ( A , D and G ). Cell proliferation was assessed at time = 0, 24, 48, and 72 h using the BrdU cell proliferation assay kit, as described in Experimental procedures . Scatter plots represent cell proliferation 0, 24, 48, and 72 h after the onset of the experiment, presented as BrdU uptake rate (n = 6). Analysis of statistical significance was performed using two-way ANOVA (for A , D , G : 0 h, F values were 0.14, 0.14, and 1.35 and p values were 0.93, 0.86, and 0.24 for concentration, time, and the interaction respectively; for A , D , G : 24 h, F values were 593.8, 1570, and 199.6 and p values were

    Article Snippet: Rabbit polyclonal anti-TRPC6 antibody (catalog number: ACC-120, epitope corresponding to amino acid residues 573–586) was from Alomone.

    Techniques: Expressing, Multiple Displacement Amplification, Transfection, Plasmid Preparation, BrdU Cell Proliferation Assay, Concentration Assay

    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.

    Journal: The Journal of Clinical Investigation

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

    doi: 10.1172/JCI129635

    Figure Lengend 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.

    Article Snippet: Cells were incubated overnight in PBS-BSA 3% with or without CFTR (Thermo Fisher Scientific, USA1-935, clone CF3) or TRPC6 (Alomone ACC-120, polyclonal) primary antibodies, followed by species-specific Alexa Fluor 647 (Invitrogen, A31571) or Alexa Fluor 488 (Invitrogen, A11006) secondary antibodies, respectively.

    Techniques: 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.

    Journal: The Journal of Clinical Investigation

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

    doi: 10.1172/JCI129635

    Figure Lengend 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.

    Article Snippet: Cells were incubated overnight in PBS-BSA 3% with or without CFTR (Thermo Fisher Scientific, USA1-935, clone CF3) or TRPC6 (Alomone ACC-120, polyclonal) primary antibodies, followed by species-specific Alexa Fluor 647 (Invitrogen, A31571) or Alexa Fluor 488 (Invitrogen, A11006) secondary antibodies, respectively.

    Techniques: 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.

    Journal: The Journal of Clinical Investigation

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

    doi: 10.1172/JCI129635

    Figure Lengend 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.

    Article Snippet: Cells were incubated overnight in PBS-BSA 3% with or without CFTR (Thermo Fisher Scientific, USA1-935, clone CF3) or TRPC6 (Alomone ACC-120, polyclonal) primary antibodies, followed by species-specific Alexa Fluor 647 (Invitrogen, A31571) or Alexa Fluor 488 (Invitrogen, A11006) secondary antibodies, respectively.

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