trpv4  (Alomone Labs)


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    Structured Review

    Alomone Labs trpv4
    FD20 content in serum. In Ctrl and DU groups, after treatment with <t>TRPV4</t> agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Trpv4, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trpv4/product/Alomone Labs
    Average 95 stars, based on 37 article reviews
    Price from $9.99 to $1999.99
    trpv4 - by Bioz Stars, 2022-08
    95/100 stars

    Images

    1) Product Images from "Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers"

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    Journal: BioMed Research International

    doi: 10.1155/2022/2777882

    FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used:

    The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used:

    TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P
    Figure Legend Snippet: TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P

    Techniques Used: Diagnostic Assay, Immunohistochemistry, Expressing

    Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used: Expressing, Mouse Assay, Staining, Concentration Assay

    2) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    3) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    4) Product Images from "Translocation of TRPV4-PI3Kγ complexes to the plasma membrane drives myofibroblast transdifferentiation"

    Article Title: Translocation of TRPV4-PI3Kγ complexes to the plasma membrane drives myofibroblast transdifferentiation

    Journal: Science signaling

    doi: 10.1126/scisignal.aau1533

    Both PI3Kγ and TRPV4 are required for myofibroblast transdifferentiation. ( A ) Representative fluorescence images showing α-SMA (green) and F-actin (red) in TRPV4 KO and PI3Kγ KO mouse lung fibroblasts (MLFs) treated with empty vector (control) lentivirus (LV), TRPV4 LV, or PI3Kγ LV, then stimulated with TGF-β. Non-transfected WT MLF were used as a positive control. Scale bar, 100 μm. ( B ) Quantification of cells with α-SMA–positive stress fibers in (A). Data are presented as % myofibroblasts (means ± SEM). N = 3 independent experiments with at least 30 cells per condition. *P
    Figure Legend Snippet: Both PI3Kγ and TRPV4 are required for myofibroblast transdifferentiation. ( A ) Representative fluorescence images showing α-SMA (green) and F-actin (red) in TRPV4 KO and PI3Kγ KO mouse lung fibroblasts (MLFs) treated with empty vector (control) lentivirus (LV), TRPV4 LV, or PI3Kγ LV, then stimulated with TGF-β. Non-transfected WT MLF were used as a positive control. Scale bar, 100 μm. ( B ) Quantification of cells with α-SMA–positive stress fibers in (A). Data are presented as % myofibroblasts (means ± SEM). N = 3 independent experiments with at least 30 cells per condition. *P

    Techniques Used: Fluorescence, Plasmid Preparation, Transfection, Positive Control

    Both TRPV4 and PI3Kγ are necessary for profibrotic responses of myofibroblasts. ( A ) Representative immunoblots showing collagen-1 in WT, TRPV4 KO, and PI3Kγ KO mouse lung fibroblasts (MLFs) treated ±TGF-β. Black lines indicate non-contiguous lanes from the same blot. GAPDH is a loading control. ( B ) Quantification of collagen-1:GAPDH band density ratios (means ± SEM) from (A). N=3 independent experiments. *P
    Figure Legend Snippet: Both TRPV4 and PI3Kγ are necessary for profibrotic responses of myofibroblasts. ( A ) Representative immunoblots showing collagen-1 in WT, TRPV4 KO, and PI3Kγ KO mouse lung fibroblasts (MLFs) treated ±TGF-β. Black lines indicate non-contiguous lanes from the same blot. GAPDH is a loading control. ( B ) Quantification of collagen-1:GAPDH band density ratios (means ± SEM) from (A). N=3 independent experiments. *P

    Techniques Used: Western Blot

    TRPV4 and PI3Kγ interact directly and translocate to the plasma membrane in response to TGF-β. (A) The binding of purified PI3Kγ to purified 6-His-TRPV4 coupled to Ni-NTA beads was assessed by immunoblotting with an antibody specific for PI3Kγ. The blots were stripped and reprobed with an antibody for TRPV4. Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( B ) The direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( C ) The binding of purified 6-His-PI3Kγ coupled to Ni-NTA beads to endogenous TRPV4 from human lung fibroblast (HLF) lysates was assessed by Western blotting. Blots were stripped and reprobed with antibodies specific for PI3Kγ, GRK2 (positive control), and TRPV2 (negative control). Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( D ) PI3Kγ immunoprecipitates (IP) from plasma membrane and cytosolic fractions of HLFs treated with TGF-β as indicated were immunoblotted for PI3Kγ and TRPV4. The line between the plasma membrane and cytosolic fractions indicates non-contiguous lanes from the same blot. Total whole cell lysate of cells treated ±TGF-β (input) was immunoblotted for TRPV4 and PI3Kγ. GAPDH is a loading control. ( E ) Quantification of TRPV4 in plasma membrane and cytosolic fractions normalized to TRPV4 in whole cell lysate as in (D). Data represent means ± SEM. N=3 independent experiments. *P≤0.05 between untreated and TGF-β conditions, using ANOVA followed by Student-Newman-Keuls multiple comparisons test.
    Figure Legend Snippet: TRPV4 and PI3Kγ interact directly and translocate to the plasma membrane in response to TGF-β. (A) The binding of purified PI3Kγ to purified 6-His-TRPV4 coupled to Ni-NTA beads was assessed by immunoblotting with an antibody specific for PI3Kγ. The blots were stripped and reprobed with an antibody for TRPV4. Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( B ) The direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( C ) The binding of purified 6-His-PI3Kγ coupled to Ni-NTA beads to endogenous TRPV4 from human lung fibroblast (HLF) lysates was assessed by Western blotting. Blots were stripped and reprobed with antibodies specific for PI3Kγ, GRK2 (positive control), and TRPV2 (negative control). Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( D ) PI3Kγ immunoprecipitates (IP) from plasma membrane and cytosolic fractions of HLFs treated with TGF-β as indicated were immunoblotted for PI3Kγ and TRPV4. The line between the plasma membrane and cytosolic fractions indicates non-contiguous lanes from the same blot. Total whole cell lysate of cells treated ±TGF-β (input) was immunoblotted for TRPV4 and PI3Kγ. GAPDH is a loading control. ( E ) Quantification of TRPV4 in plasma membrane and cytosolic fractions normalized to TRPV4 in whole cell lysate as in (D). Data represent means ± SEM. N=3 independent experiments. *P≤0.05 between untreated and TGF-β conditions, using ANOVA followed by Student-Newman-Keuls multiple comparisons test.

    Techniques Used: Binding Assay, Purification, SPR Assay, Western Blot, Positive Control, Negative Control

    Schematic working model showing TGF-β–stimulated activation of TRPV4-PI3Kγ complexes in myofibroblast transdifferentiation. Our data suggest that TRPV4 binds to the non-catalytic, N-terminal domain of PI3Kγ, forming a TRPV4-PI3Kγ complex. Upon TGF-β stimulation, the N-terminus of PI3Kγ acts as a scaffold, recruiting TRPV4-PI3Kγ complexes from the cytoplasm to the plasma membrane. This translocation leads to an increase in TRPV4-mediated Ca 2+ influx, which promotes myofibroblast transdifferentiation.
    Figure Legend Snippet: Schematic working model showing TGF-β–stimulated activation of TRPV4-PI3Kγ complexes in myofibroblast transdifferentiation. Our data suggest that TRPV4 binds to the non-catalytic, N-terminal domain of PI3Kγ, forming a TRPV4-PI3Kγ complex. Upon TGF-β stimulation, the N-terminus of PI3Kγ acts as a scaffold, recruiting TRPV4-PI3Kγ complexes from the cytoplasm to the plasma membrane. This translocation leads to an increase in TRPV4-mediated Ca 2+ influx, which promotes myofibroblast transdifferentiation.

    Techniques Used: Activation Assay, Translocation Assay

    TRPV4 selectively mediates TGF-β–driven PI3Kγ activity. ( A ) Representative Western blot showing TRPV4, phosphorylated AKT [p-AKT (Ser 473 )], total AKT (t-AKT), and GAPDH (loading control) in lysates of human lung fibroblast (HLF) treated with TRPV4 or scrambled siRNA ±TGF-β as indicated. ( B ) Quantification of p-AKT (Ser 473 ) relative to total AKT as in (A) by densitometry. *P
    Figure Legend Snippet: TRPV4 selectively mediates TGF-β–driven PI3Kγ activity. ( A ) Representative Western blot showing TRPV4, phosphorylated AKT [p-AKT (Ser 473 )], total AKT (t-AKT), and GAPDH (loading control) in lysates of human lung fibroblast (HLF) treated with TRPV4 or scrambled siRNA ±TGF-β as indicated. ( B ) Quantification of p-AKT (Ser 473 ) relative to total AKT as in (A) by densitometry. *P

    Techniques Used: Activity Assay, Western Blot

    TRPV4 and PI3Kγ are mutually required for translocation to the plasma membrane. (A ) Representative confocal images showing TRPV4 in wild-type (WT) and PI3Kγ knockout (KO) murine lung fibroblasts (MLFs) ±TGF-β as indicated and the corresponding plot profiles of the TRPV4 immunofluorescence. Orange lines indicate regions where plot profiles were obtained. White boxes indicate areas shown in higher magnification in insets. ( B ) Quantification of plasma membrane:cytoplasm fluorescence for experiments in (A). N = 3 independent experiments with at least 30 cells per condition. ***P
    Figure Legend Snippet: TRPV4 and PI3Kγ are mutually required for translocation to the plasma membrane. (A ) Representative confocal images showing TRPV4 in wild-type (WT) and PI3Kγ knockout (KO) murine lung fibroblasts (MLFs) ±TGF-β as indicated and the corresponding plot profiles of the TRPV4 immunofluorescence. Orange lines indicate regions where plot profiles were obtained. White boxes indicate areas shown in higher magnification in insets. ( B ) Quantification of plasma membrane:cytoplasm fluorescence for experiments in (A). N = 3 independent experiments with at least 30 cells per condition. ***P

    Techniques Used: Translocation Assay, Knock-Out, Immunofluorescence, Fluorescence

    PI3Kγ is required for TRPV4 ion channel function. ( A ) Representative plots showing the effects of scrambled, PI3Kα, and PI3Kγ siRNA on Ca 2+ influx induced by the TRPV4 agonist GSK1016790A (GSK) in human lung fibroblasts. Ca 2+ influx was measured in relative fluorescence units (RFU) using Calcium 5 dye on intact human lung fibroblast (19Lu) monolayers treated with scrambled, PI3Kα, or PI3Kγ siRNA. ( B ) Quantification of experiments in (A). ***P
    Figure Legend Snippet: PI3Kγ is required for TRPV4 ion channel function. ( A ) Representative plots showing the effects of scrambled, PI3Kα, and PI3Kγ siRNA on Ca 2+ influx induced by the TRPV4 agonist GSK1016790A (GSK) in human lung fibroblasts. Ca 2+ influx was measured in relative fluorescence units (RFU) using Calcium 5 dye on intact human lung fibroblast (19Lu) monolayers treated with scrambled, PI3Kα, or PI3Kγ siRNA. ( B ) Quantification of experiments in (A). ***P

    Techniques Used: Fluorescence

    The non-catalytic, amino-terminal domain of PI3Kγ is necessary and sufficient for TRPV4 translocation and myofibroblast transdifferentiation. ( A ) Domain structure of WT and mutant PI3Kγ constructs consisting of only the non-catalytic N-terminal domain (N-term) or lacking both the N-terminal domain and the ATP binding site in the catalytic domain (N-del). ( B ) Immunoblotting of His-tagged N-del or N-term forms of PI3Kγ coupled to Ni-NTA beads incubated with lysates of human lung fibroblasts (HLFs). Blots were probed with an antibody recognizing TRPV4, then stripped and reprobed for PI3Kγ. N=3 independent experiments. ( C ) Direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-N-term-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( D ) Representative fluorescence images showing TRPV4 in PI3Kγ KO murine lung fibroblasts (MLFs) transfected with N-del or N-term PI3Kγ lentivirus (LV) and treated with TGF-β and the corresponding plot profiles of TRPV4 immunofluorescence. White arrows indicate TRPV4 at the plasma membrane; orange lines indicate regions where plot profiles were obtained; white boxes indicate higher magnification insets. Scale bar, 50 μm. ( E ) Quantification of results from (D). N = 3 independent experiments with at least 30 cells per condition. ***P
    Figure Legend Snippet: The non-catalytic, amino-terminal domain of PI3Kγ is necessary and sufficient for TRPV4 translocation and myofibroblast transdifferentiation. ( A ) Domain structure of WT and mutant PI3Kγ constructs consisting of only the non-catalytic N-terminal domain (N-term) or lacking both the N-terminal domain and the ATP binding site in the catalytic domain (N-del). ( B ) Immunoblotting of His-tagged N-del or N-term forms of PI3Kγ coupled to Ni-NTA beads incubated with lysates of human lung fibroblasts (HLFs). Blots were probed with an antibody recognizing TRPV4, then stripped and reprobed for PI3Kγ. N=3 independent experiments. ( C ) Direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-N-term-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( D ) Representative fluorescence images showing TRPV4 in PI3Kγ KO murine lung fibroblasts (MLFs) transfected with N-del or N-term PI3Kγ lentivirus (LV) and treated with TGF-β and the corresponding plot profiles of TRPV4 immunofluorescence. White arrows indicate TRPV4 at the plasma membrane; orange lines indicate regions where plot profiles were obtained; white boxes indicate higher magnification insets. Scale bar, 50 μm. ( E ) Quantification of results from (D). N = 3 independent experiments with at least 30 cells per condition. ***P

    Techniques Used: Translocation Assay, Mutagenesis, Construct, Binding Assay, Incubation, Purification, SPR Assay, Fluorescence, Transfection, Immunofluorescence

    5) Product Images from "Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)"

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    Journal: bioRxiv

    doi: 10.1101/2020.02.10.941732

    Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P
    Figure Legend Snippet: Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P

    Techniques Used: Staining, Expressing, Fluorescence

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.
    Figure Legend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Techniques Used: Expressing, Western Blot, Blocking Assay

    6) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    7) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    8) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    9) Product Images from "Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension"

    Article Title: Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

    Journal: Journal of Lipid Research

    doi: 10.1016/j.jlr.2021.100145

    Most membrane TRPV4 does not partition into raft domains or interact with caveolar proteins. A: Western blot; detergent-free lipid raft isolation in primary TM cells. The supernatant fraction contains cytosolic proteins; fractions 1 and 2 (5% sucrose) contain nonlipid-raft membrane proteins, fractions 3–6 (5–35% sucrose) contain lipid raft membranes, and fractions 7 and 8 (pellet, 45% sucrose) contain unsuspended proteins and cell nuclei. TRPV4 protein is predominantly confined to fraction 2; fraction 4 is associated with flotillin-1 (48 kDa) and Cav-1 (22 kDa), whereas the supernatant/cytosol fraction associates with α-SMA (42 kDa). B: Coimmunoprecipitation. TRPV4-Cav-1 interaction assessed with the Cav-1 antibody for TRPV4 pulldown in control, MβCD, and MβCD:cholesterol-treated samples. Input bands, whole-cell lysate; bound bands, Cav-1-bound protein fraction; unbound bands, flow-through fractions. Cav-1-bound fractions show modest precipitation of the ∼75 kDa TRPV4 isoform. The absence of Cav-1 expression in unbound fractions confirms the quantitative immunoprecipitation of Cav-1. C, D: Immunohistochemistry, for control, MβCD, and MβCD:cholesterol-treated cells. C: Double immunolabeling for TRPV4 and Cav-1. TRPV4-ir (red arrowheads) does not colocalize with Cav1-ir puncta (green). D: Double immunolabeling for TRPV4 and flotillin. TRPV4-ir (red puncta) does not colocalize with flotillin-ir puncta (green). The inset is shown at higher magnification insets as supplemental Fig. S3 . The scale bar represents 20 μm.
    Figure Legend Snippet: Most membrane TRPV4 does not partition into raft domains or interact with caveolar proteins. A: Western blot; detergent-free lipid raft isolation in primary TM cells. The supernatant fraction contains cytosolic proteins; fractions 1 and 2 (5% sucrose) contain nonlipid-raft membrane proteins, fractions 3–6 (5–35% sucrose) contain lipid raft membranes, and fractions 7 and 8 (pellet, 45% sucrose) contain unsuspended proteins and cell nuclei. TRPV4 protein is predominantly confined to fraction 2; fraction 4 is associated with flotillin-1 (48 kDa) and Cav-1 (22 kDa), whereas the supernatant/cytosol fraction associates with α-SMA (42 kDa). B: Coimmunoprecipitation. TRPV4-Cav-1 interaction assessed with the Cav-1 antibody for TRPV4 pulldown in control, MβCD, and MβCD:cholesterol-treated samples. Input bands, whole-cell lysate; bound bands, Cav-1-bound protein fraction; unbound bands, flow-through fractions. Cav-1-bound fractions show modest precipitation of the ∼75 kDa TRPV4 isoform. The absence of Cav-1 expression in unbound fractions confirms the quantitative immunoprecipitation of Cav-1. C, D: Immunohistochemistry, for control, MβCD, and MβCD:cholesterol-treated cells. C: Double immunolabeling for TRPV4 and Cav-1. TRPV4-ir (red arrowheads) does not colocalize with Cav1-ir puncta (green). D: Double immunolabeling for TRPV4 and flotillin. TRPV4-ir (red puncta) does not colocalize with flotillin-ir puncta (green). The inset is shown at higher magnification insets as supplemental Fig. S3 . The scale bar represents 20 μm.

    Techniques Used: Western Blot, Isolation, Expressing, Immunoprecipitation, Immunohistochemistry, Immunolabeling

    Cholesterol depletion enhances TRPV4-mediated membrane currents in Xenopus oocytes. A: Representative current traces from TRPV4-expressing oocytes and uninjected control oocytes in control solution or after 45 min exposure to 50 μM MβCD. B: I/V curves of TRPV4-expressing oocytes exposed to control solution or MβCD, with uninjected oocytes in inset. Summarized currents obtained at −85 mV are shown in the lower inset. The magnitude of TRPV4-mediated currents (at V m = −85 mV) was compared using Student's t -test. ∗∗ P
    Figure Legend Snippet: Cholesterol depletion enhances TRPV4-mediated membrane currents in Xenopus oocytes. A: Representative current traces from TRPV4-expressing oocytes and uninjected control oocytes in control solution or after 45 min exposure to 50 μM MβCD. B: I/V curves of TRPV4-expressing oocytes exposed to control solution or MβCD, with uninjected oocytes in inset. Summarized currents obtained at −85 mV are shown in the lower inset. The magnitude of TRPV4-mediated currents (at V m = −85 mV) was compared using Student's t -test. ∗∗ P

    Techniques Used: Expressing

    Cholesterol depletion increases the number of TRPV4-ir puncta. TRPV4 immunolabeling of primary TM cells. Representative examples of (A) control, (B) 1 h treatment with MβCD, and (C) 1 h treatment with MβCD:cholesterol. Inset: zoomed-in region with TRPV4-ir puncta (arrow). D: Summary of three independent experiments, normalized for control cells. The number of TRPV4 puncta is upregulated after incubation with MβCD. ∗ P
    Figure Legend Snippet: Cholesterol depletion increases the number of TRPV4-ir puncta. TRPV4 immunolabeling of primary TM cells. Representative examples of (A) control, (B) 1 h treatment with MβCD, and (C) 1 h treatment with MβCD:cholesterol. Inset: zoomed-in region with TRPV4-ir puncta (arrow). D: Summary of three independent experiments, normalized for control cells. The number of TRPV4 puncta is upregulated after incubation with MβCD. ∗ P

    Techniques Used: Immunolabeling, Incubation

    Cholesterol depletion increases the amplitude of TRPV4 agonist-induced Ca 2+ signals. A, B: Ratiometric signals in Fura-2 AM-loaded cells. A: GSK101-induced elevations are increased in MβCD-treated cells (n = 8–10). B: Averaged data. MβCD (blue bar) augmented, whereas MβCD:cholesterol (green bar) reduced the amplitude of agonist-induced fluorescence. C: Fluorimetry, cell populations in 96 wells. About 5 nM GSK101 increased Fluo-4 fluorescence. Its effect was facilitated (∼12%) by MβCD (N = 4). ∗∗ P
    Figure Legend Snippet: Cholesterol depletion increases the amplitude of TRPV4 agonist-induced Ca 2+ signals. A, B: Ratiometric signals in Fura-2 AM-loaded cells. A: GSK101-induced elevations are increased in MβCD-treated cells (n = 8–10). B: Averaged data. MβCD (blue bar) augmented, whereas MβCD:cholesterol (green bar) reduced the amplitude of agonist-induced fluorescence. C: Fluorimetry, cell populations in 96 wells. About 5 nM GSK101 increased Fluo-4 fluorescence. Its effect was facilitated (∼12%) by MβCD (N = 4). ∗∗ P

    Techniques Used: Fluorescence

    10) Product Images from "Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers"

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    Journal: BioMed Research International

    doi: 10.1155/2022/2777882

    FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used:

    The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used:

    TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P
    Figure Legend Snippet: TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P

    Techniques Used: Diagnostic Assay, Immunohistochemistry, Expressing

    Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Figure Legend Snippet: Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Techniques Used: Expressing, Mouse Assay, Staining, Concentration Assay

    11) Product Images from "Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)"

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    Journal: bioRxiv

    doi: 10.1101/2020.02.10.941732

    Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P
    Figure Legend Snippet: Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P

    Techniques Used: Staining, Expressing, Fluorescence

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.
    Figure Legend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Techniques Used: Expressing, Western Blot, Blocking Assay

    12) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    13) Product Images from "TRPV4 and AQP4 Channels Synergistically Regulate Cell Volume and Calcium Homeostasis in Retinal Müller Glia"

    Article Title: TRPV4 and AQP4 Channels Synergistically Regulate Cell Volume and Calcium Homeostasis in Retinal Müller Glia

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1987-15.2015

    Proposed schema of TRPV4–AQP4 interactions in the Müller cell end foot. Hypotonic stress stimulates fluxes of water after the osmotic gradient. The resulting increase in cell volume stretches the plasma membrane, activating TRPV4 and a
    Figure Legend Snippet: Proposed schema of TRPV4–AQP4 interactions in the Müller cell end foot. Hypotonic stress stimulates fluxes of water after the osmotic gradient. The resulting increase in cell volume stretches the plasma membrane, activating TRPV4 and a

    Techniques Used:

    TRPV4 and AQP4 colocalize in the mouse inner retina, yet TRPV4 and AQP4 trafficking are independent of each other. Ai , Vertical cryosections of WT mouse retinas immunolabeled for TRPV4 (Alexa Fluor 488) and AQP4 (Alexa Fluor 594) show preferential colocalization
    Figure Legend Snippet: TRPV4 and AQP4 colocalize in the mouse inner retina, yet TRPV4 and AQP4 trafficking are independent of each other. Ai , Vertical cryosections of WT mouse retinas immunolabeled for TRPV4 (Alexa Fluor 488) and AQP4 (Alexa Fluor 594) show preferential colocalization

    Techniques Used: Immunolabeling

    TRPV4 blockers suppress the swelling of Aqp4 −/− Müller glia loaded with calcein AM. A , The TRPV4 antagonist HC-06 (1 μ m ) suppresses HTS-induced increases in Aqp4 −/− glial volume (190 mOsm; n = 9 cells from
    Figure Legend Snippet: TRPV4 blockers suppress the swelling of Aqp4 −/− Müller glia loaded with calcein AM. A , The TRPV4 antagonist HC-06 (1 μ m ) suppresses HTS-induced increases in Aqp4 −/− glial volume (190 mOsm; n = 9 cells from

    Techniques Used:

    TRPV4-induced Ca 2+ entry regulates Müller glial swelling and RVD. A , Cell volume changes in a fura-5F-loaded Müller cell. The sum of 340/380 emissions, adjusted to yield calcium-insensitive intensity values, shows dose-dependent decreases
    Figure Legend Snippet: TRPV4-induced Ca 2+ entry regulates Müller glial swelling and RVD. A , Cell volume changes in a fura-5F-loaded Müller cell. The sum of 340/380 emissions, adjusted to yield calcium-insensitive intensity values, shows dose-dependent decreases

    Techniques Used:

    Expression of Trpv4 , Aqp4 , and Kir4.1 transcripts is effected by genetic elimination of TRPV4 or AQP4. Semiquantitative RT-PCR analysis of mRNA levels in WT, Trpv4 −/− , and Aqp4 −/− mouse retinas. Fold change in mRNA expression
    Figure Legend Snippet: Expression of Trpv4 , Aqp4 , and Kir4.1 transcripts is effected by genetic elimination of TRPV4 or AQP4. Semiquantitative RT-PCR analysis of mRNA levels in WT, Trpv4 −/− , and Aqp4 −/− mouse retinas. Fold change in mRNA expression

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    AQP4 is not required for agonist-induced TRPV4 activation. A , Averaged I–V curves of GSK101-induced currents in WT (black trace) and Aqp4 −/− (blue trace) Müller cells. B , Representative time courses of the GSK101-evoked
    Figure Legend Snippet: AQP4 is not required for agonist-induced TRPV4 activation. A , Averaged I–V curves of GSK101-induced currents in WT (black trace) and Aqp4 −/− (blue trace) Müller cells. B , Representative time courses of the GSK101-evoked

    Techniques Used: Activation Assay

    AQP4 amplifies glial HTS-induced TRPV4 activation with moderate (140 mOsm) but not extreme (90 mOsm) HTS. A , B , Ca 2+ signals in response to 140 mOsm HTS were blocked in Trpv4 −/− ( n = 50) and Aqp4 −/− ( n = 64) Müller
    Figure Legend Snippet: AQP4 amplifies glial HTS-induced TRPV4 activation with moderate (140 mOsm) but not extreme (90 mOsm) HTS. A , B , Ca 2+ signals in response to 140 mOsm HTS were blocked in Trpv4 −/− ( n = 50) and Aqp4 −/− ( n = 64) Müller

    Techniques Used: Activation Assay

    AQP4 channels amplify stretch-induced TRPV4 activation. A , Oocytes expressing TRPV4 and AQP4 alone or in combination were voltage clamped at V m = −30 mV and exposed to 100 n m GSK101 (indicated by black bar). After 60 s GSK exposure, an I–V
    Figure Legend Snippet: AQP4 channels amplify stretch-induced TRPV4 activation. A , Oocytes expressing TRPV4 and AQP4 alone or in combination were voltage clamped at V m = −30 mV and exposed to 100 n m GSK101 (indicated by black bar). After 60 s GSK exposure, an I–V

    Techniques Used: Activation Assay, Expressing

    AQP4 regulates the amplitude and kinetics of the swelling-induced TRPV4-mediated transmembrane current. A , B , Representative time course of the HTS (235 mOsm)-evoked whole-cell current in WT (black traces) and Aqp4 −/− (blue traces) Müller
    Figure Legend Snippet: AQP4 regulates the amplitude and kinetics of the swelling-induced TRPV4-mediated transmembrane current. A , B , Representative time course of the HTS (235 mOsm)-evoked whole-cell current in WT (black traces) and Aqp4 −/− (blue traces) Müller

    Techniques Used:

    14) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    15) Product Images from "Extracellular Vesicles From Pathological Microenvironment Induce Endothelial Cell Transformation and Abnormal Angiogenesis via Modulation of TRPV4 Channels"

    Article Title: Extracellular Vesicles From Pathological Microenvironment Induce Endothelial Cell Transformation and Abnormal Angiogenesis via Modulation of TRPV4 Channels

    Journal: Frontiers in Cell and Developmental Biology

    doi: 10.3389/fcell.2019.00344

    TCM exposure downregulates functional expression of TRPV4 channels in hTEC. (A) Western blot analysis showing decreased expression of TRPV4 protein in hTEC when compared to hNEC. (B) Quantitative analysis of western blots revealed significant ( ∗ p ≤ 0.05) reduction of TRPV4 expression in hTEC. (C) Average traces showing calcium influx in response to the TRPV4 agonist, GSK1016790A (100 nM), in Fluo-4 loaded hNEC and hTEC. Arrow indicates the time of stimulation with TRPV4 agonist. (D) Quantitative analysis of calcium influx showed significant **** p ≤ 0.0001 reduction in TRPV4-mediated calcium influx in hTEC compared to hNEC (F/F0 = ratio of normalized fluorescent intensity relative to time 0). The results shown are a mean ± SEM from three independent experiments.
    Figure Legend Snippet: TCM exposure downregulates functional expression of TRPV4 channels in hTEC. (A) Western blot analysis showing decreased expression of TRPV4 protein in hTEC when compared to hNEC. (B) Quantitative analysis of western blots revealed significant ( ∗ p ≤ 0.05) reduction of TRPV4 expression in hTEC. (C) Average traces showing calcium influx in response to the TRPV4 agonist, GSK1016790A (100 nM), in Fluo-4 loaded hNEC and hTEC. Arrow indicates the time of stimulation with TRPV4 agonist. (D) Quantitative analysis of calcium influx showed significant **** p ≤ 0.0001 reduction in TRPV4-mediated calcium influx in hTEC compared to hNEC (F/F0 = ratio of normalized fluorescent intensity relative to time 0). The results shown are a mean ± SEM from three independent experiments.

    Techniques Used: Functional Assay, Expressing, Western Blot

    16) Product Images from "Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)"

    Article Title: Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos)

    Journal: bioRxiv

    doi: 10.1101/2020.02.10.941732

    Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.
    Figure Legend Snippet: Microscopic images showing localization of TRPV4 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV4 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV4 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the presence of TRPV4 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Techniques Used: Expressing

    Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P
    Figure Legend Snippet: Prevalence of physiologically relevant thermosensitive TRPV channels in mature duck sperm. Flow cytometric evaluation of duck sperm stained for physiologically relevant thermosensitive TRPV channels are shown. A. Representative dot-plots showing percentage of cells expressing TRPV1, TRPV3, TRPV4 channels detected by Ab-1 (antibodies from Alomone labs) antibody specific for each TRPV channel. B. Histograms showing percentage of cells expressing TRPV channels and corresponding Mean Fluorescence Intensity (MFI) of TRPV channels detected by Ab2 antibody of each channel (from Sigma Aldrich), expressed as fold change in comparison to MFI of unstained cells. n = 3, unpaired T-test. ** = P

    Techniques Used: Staining, Expressing, Fluorescence

    Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.
    Figure Legend Snippet: Endogenous expression of TRPV channels in duck sperm. Western blot analysis of duck sperm extracts probed with different TRPV-specific antibodies are shown. A. TRPV1 specific band is detected by a specific antibody (directed against the C-terminus of TRPV1, Alomone Labs) in absence but not in presence of its blocking peptide; B. Western blot analysis with antibody that detects TRPV2 (raised against the C-terminus, Alomone Labs). C. Two different antibodies detecting TRPV3 [raised against the C-terminus, (Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect similar expression pattern of TRPV3. D. Two different antibodies raised against the TRPV4 [raised against C-terminus, Ab1: Alomone Labs) and N-terminus (Ab3: Sigma Aldrich)] detect TRPV4 at the expected size. E. Two different antibodies raised against the C-terminus of TRPV5 (Ab1: Alomone Labs and Ab2: Sigma-Aldrich) detects TRPV5 at expected size. F. A specific antibody raised against the C-terminus of TRPV6 (Ab-1: Alomone Labs) detects TRPV6 in absence but not in presence of its blocking peptide.

    Techniques Used: Expressing, Western Blot, Blocking Assay

    17) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    18) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    19) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    20) Product Images from "Translocation of TRPV4-PI3Kγ complexes to the plasma membrane drives myofibroblast transdifferentiation"

    Article Title: Translocation of TRPV4-PI3Kγ complexes to the plasma membrane drives myofibroblast transdifferentiation

    Journal: Science signaling

    doi: 10.1126/scisignal.aau1533

    Both PI3Kγ and TRPV4 are required for myofibroblast transdifferentiation. ( A ) Representative fluorescence images showing α-SMA (green) and F-actin (red) in TRPV4 KO and PI3Kγ KO mouse lung fibroblasts (MLFs) treated with empty vector (control) lentivirus (LV), TRPV4 LV, or PI3Kγ LV, then stimulated with TGF-β. Non-transfected WT MLF were used as a positive control. Scale bar, 100 μm. ( B ) Quantification of cells with α-SMA–positive stress fibers in (A). Data are presented as % myofibroblasts (means ± SEM). N = 3 independent experiments with at least 30 cells per condition. *P
    Figure Legend Snippet: Both PI3Kγ and TRPV4 are required for myofibroblast transdifferentiation. ( A ) Representative fluorescence images showing α-SMA (green) and F-actin (red) in TRPV4 KO and PI3Kγ KO mouse lung fibroblasts (MLFs) treated with empty vector (control) lentivirus (LV), TRPV4 LV, or PI3Kγ LV, then stimulated with TGF-β. Non-transfected WT MLF were used as a positive control. Scale bar, 100 μm. ( B ) Quantification of cells with α-SMA–positive stress fibers in (A). Data are presented as % myofibroblasts (means ± SEM). N = 3 independent experiments with at least 30 cells per condition. *P

    Techniques Used: Fluorescence, Plasmid Preparation, Transfection, Positive Control

    Both TRPV4 and PI3Kγ are necessary for profibrotic responses of myofibroblasts. ( A ) Representative immunoblots showing collagen-1 in WT, TRPV4 KO, and PI3Kγ KO mouse lung fibroblasts (MLFs) treated ±TGF-β. Black lines indicate non-contiguous lanes from the same blot. GAPDH is a loading control. ( B ) Quantification of collagen-1:GAPDH band density ratios (means ± SEM) from (A). N=3 independent experiments. *P
    Figure Legend Snippet: Both TRPV4 and PI3Kγ are necessary for profibrotic responses of myofibroblasts. ( A ) Representative immunoblots showing collagen-1 in WT, TRPV4 KO, and PI3Kγ KO mouse lung fibroblasts (MLFs) treated ±TGF-β. Black lines indicate non-contiguous lanes from the same blot. GAPDH is a loading control. ( B ) Quantification of collagen-1:GAPDH band density ratios (means ± SEM) from (A). N=3 independent experiments. *P

    Techniques Used: Western Blot

    TRPV4 and PI3Kγ interact directly and translocate to the plasma membrane in response to TGF-β. (A) The binding of purified PI3Kγ to purified 6-His-TRPV4 coupled to Ni-NTA beads was assessed by immunoblotting with an antibody specific for PI3Kγ. The blots were stripped and reprobed with an antibody for TRPV4. Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( B ) The direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( C ) The binding of purified 6-His-PI3Kγ coupled to Ni-NTA beads to endogenous TRPV4 from human lung fibroblast (HLF) lysates was assessed by Western blotting. Blots were stripped and reprobed with antibodies specific for PI3Kγ, GRK2 (positive control), and TRPV2 (negative control). Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( D ) PI3Kγ immunoprecipitates (IP) from plasma membrane and cytosolic fractions of HLFs treated with TGF-β as indicated were immunoblotted for PI3Kγ and TRPV4. The line between the plasma membrane and cytosolic fractions indicates non-contiguous lanes from the same blot. Total whole cell lysate of cells treated ±TGF-β (input) was immunoblotted for TRPV4 and PI3Kγ. GAPDH is a loading control. ( E ) Quantification of TRPV4 in plasma membrane and cytosolic fractions normalized to TRPV4 in whole cell lysate as in (D). Data represent means ± SEM. N=3 independent experiments. *P≤0.05 between untreated and TGF-β conditions, using ANOVA followed by Student-Newman-Keuls multiple comparisons test.
    Figure Legend Snippet: TRPV4 and PI3Kγ interact directly and translocate to the plasma membrane in response to TGF-β. (A) The binding of purified PI3Kγ to purified 6-His-TRPV4 coupled to Ni-NTA beads was assessed by immunoblotting with an antibody specific for PI3Kγ. The blots were stripped and reprobed with an antibody for TRPV4. Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( B ) The direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( C ) The binding of purified 6-His-PI3Kγ coupled to Ni-NTA beads to endogenous TRPV4 from human lung fibroblast (HLF) lysates was assessed by Western blotting. Blots were stripped and reprobed with antibodies specific for PI3Kγ, GRK2 (positive control), and TRPV2 (negative control). Control indicates beads alone (no protein bound to beads), N=3 independent experiments. ( D ) PI3Kγ immunoprecipitates (IP) from plasma membrane and cytosolic fractions of HLFs treated with TGF-β as indicated were immunoblotted for PI3Kγ and TRPV4. The line between the plasma membrane and cytosolic fractions indicates non-contiguous lanes from the same blot. Total whole cell lysate of cells treated ±TGF-β (input) was immunoblotted for TRPV4 and PI3Kγ. GAPDH is a loading control. ( E ) Quantification of TRPV4 in plasma membrane and cytosolic fractions normalized to TRPV4 in whole cell lysate as in (D). Data represent means ± SEM. N=3 independent experiments. *P≤0.05 between untreated and TGF-β conditions, using ANOVA followed by Student-Newman-Keuls multiple comparisons test.

    Techniques Used: Binding Assay, Purification, SPR Assay, Western Blot, Positive Control, Negative Control

    Schematic working model showing TGF-β–stimulated activation of TRPV4-PI3Kγ complexes in myofibroblast transdifferentiation. Our data suggest that TRPV4 binds to the non-catalytic, N-terminal domain of PI3Kγ, forming a TRPV4-PI3Kγ complex. Upon TGF-β stimulation, the N-terminus of PI3Kγ acts as a scaffold, recruiting TRPV4-PI3Kγ complexes from the cytoplasm to the plasma membrane. This translocation leads to an increase in TRPV4-mediated Ca 2+ influx, which promotes myofibroblast transdifferentiation.
    Figure Legend Snippet: Schematic working model showing TGF-β–stimulated activation of TRPV4-PI3Kγ complexes in myofibroblast transdifferentiation. Our data suggest that TRPV4 binds to the non-catalytic, N-terminal domain of PI3Kγ, forming a TRPV4-PI3Kγ complex. Upon TGF-β stimulation, the N-terminus of PI3Kγ acts as a scaffold, recruiting TRPV4-PI3Kγ complexes from the cytoplasm to the plasma membrane. This translocation leads to an increase in TRPV4-mediated Ca 2+ influx, which promotes myofibroblast transdifferentiation.

    Techniques Used: Activation Assay, Translocation Assay

    TRPV4 selectively mediates TGF-β–driven PI3Kγ activity. ( A ) Representative Western blot showing TRPV4, phosphorylated AKT [p-AKT (Ser 473 )], total AKT (t-AKT), and GAPDH (loading control) in lysates of human lung fibroblast (HLF) treated with TRPV4 or scrambled siRNA ±TGF-β as indicated. ( B ) Quantification of p-AKT (Ser 473 ) relative to total AKT as in (A) by densitometry. *P
    Figure Legend Snippet: TRPV4 selectively mediates TGF-β–driven PI3Kγ activity. ( A ) Representative Western blot showing TRPV4, phosphorylated AKT [p-AKT (Ser 473 )], total AKT (t-AKT), and GAPDH (loading control) in lysates of human lung fibroblast (HLF) treated with TRPV4 or scrambled siRNA ±TGF-β as indicated. ( B ) Quantification of p-AKT (Ser 473 ) relative to total AKT as in (A) by densitometry. *P

    Techniques Used: Activity Assay, Western Blot

    TRPV4 and PI3Kγ are mutually required for translocation to the plasma membrane. (A ) Representative confocal images showing TRPV4 in wild-type (WT) and PI3Kγ knockout (KO) murine lung fibroblasts (MLFs) ±TGF-β as indicated and the corresponding plot profiles of the TRPV4 immunofluorescence. Orange lines indicate regions where plot profiles were obtained. White boxes indicate areas shown in higher magnification in insets. ( B ) Quantification of plasma membrane:cytoplasm fluorescence for experiments in (A). N = 3 independent experiments with at least 30 cells per condition. ***P
    Figure Legend Snippet: TRPV4 and PI3Kγ are mutually required for translocation to the plasma membrane. (A ) Representative confocal images showing TRPV4 in wild-type (WT) and PI3Kγ knockout (KO) murine lung fibroblasts (MLFs) ±TGF-β as indicated and the corresponding plot profiles of the TRPV4 immunofluorescence. Orange lines indicate regions where plot profiles were obtained. White boxes indicate areas shown in higher magnification in insets. ( B ) Quantification of plasma membrane:cytoplasm fluorescence for experiments in (A). N = 3 independent experiments with at least 30 cells per condition. ***P

    Techniques Used: Translocation Assay, Knock-Out, Immunofluorescence, Fluorescence

    PI3Kγ is required for TRPV4 ion channel function. ( A ) Representative plots showing the effects of scrambled, PI3Kα, and PI3Kγ siRNA on Ca 2+ influx induced by the TRPV4 agonist GSK1016790A (GSK) in human lung fibroblasts. Ca 2+ influx was measured in relative fluorescence units (RFU) using Calcium 5 dye on intact human lung fibroblast (19Lu) monolayers treated with scrambled, PI3Kα, or PI3Kγ siRNA. ( B ) Quantification of experiments in (A). ***P
    Figure Legend Snippet: PI3Kγ is required for TRPV4 ion channel function. ( A ) Representative plots showing the effects of scrambled, PI3Kα, and PI3Kγ siRNA on Ca 2+ influx induced by the TRPV4 agonist GSK1016790A (GSK) in human lung fibroblasts. Ca 2+ influx was measured in relative fluorescence units (RFU) using Calcium 5 dye on intact human lung fibroblast (19Lu) monolayers treated with scrambled, PI3Kα, or PI3Kγ siRNA. ( B ) Quantification of experiments in (A). ***P

    Techniques Used: Fluorescence

    The non-catalytic, amino-terminal domain of PI3Kγ is necessary and sufficient for TRPV4 translocation and myofibroblast transdifferentiation. ( A ) Domain structure of WT and mutant PI3Kγ constructs consisting of only the non-catalytic N-terminal domain (N-term) or lacking both the N-terminal domain and the ATP binding site in the catalytic domain (N-del). ( B ) Immunoblotting of His-tagged N-del or N-term forms of PI3Kγ coupled to Ni-NTA beads incubated with lysates of human lung fibroblasts (HLFs). Blots were probed with an antibody recognizing TRPV4, then stripped and reprobed for PI3Kγ. N=3 independent experiments. ( C ) Direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-N-term-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( D ) Representative fluorescence images showing TRPV4 in PI3Kγ KO murine lung fibroblasts (MLFs) transfected with N-del or N-term PI3Kγ lentivirus (LV) and treated with TGF-β and the corresponding plot profiles of TRPV4 immunofluorescence. White arrows indicate TRPV4 at the plasma membrane; orange lines indicate regions where plot profiles were obtained; white boxes indicate higher magnification insets. Scale bar, 50 μm. ( E ) Quantification of results from (D). N = 3 independent experiments with at least 30 cells per condition. ***P
    Figure Legend Snippet: The non-catalytic, amino-terminal domain of PI3Kγ is necessary and sufficient for TRPV4 translocation and myofibroblast transdifferentiation. ( A ) Domain structure of WT and mutant PI3Kγ constructs consisting of only the non-catalytic N-terminal domain (N-term) or lacking both the N-terminal domain and the ATP binding site in the catalytic domain (N-del). ( B ) Immunoblotting of His-tagged N-del or N-term forms of PI3Kγ coupled to Ni-NTA beads incubated with lysates of human lung fibroblasts (HLFs). Blots were probed with an antibody recognizing TRPV4, then stripped and reprobed for PI3Kγ. N=3 independent experiments. ( C ) Direct interaction between immobilized purified 6-His-TRPV4 and purified 6-His-N-term-PI3Kγ was measured by surface plasmon resonance. N=3 independent experiments. ( D ) Representative fluorescence images showing TRPV4 in PI3Kγ KO murine lung fibroblasts (MLFs) transfected with N-del or N-term PI3Kγ lentivirus (LV) and treated with TGF-β and the corresponding plot profiles of TRPV4 immunofluorescence. White arrows indicate TRPV4 at the plasma membrane; orange lines indicate regions where plot profiles were obtained; white boxes indicate higher magnification insets. Scale bar, 50 μm. ( E ) Quantification of results from (D). N = 3 independent experiments with at least 30 cells per condition. ***P

    Techniques Used: Translocation Assay, Mutagenesis, Construct, Binding Assay, Incubation, Purification, SPR Assay, Fluorescence, Transfection, Immunofluorescence

    21) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    22) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    23) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    24) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    25) Product Images from "Activation of Trpv4 reduces the hyperproliferative phenotype of cystic cholangiocytes from an animal model of ARPKD"

    Article Title: Activation of Trpv4 reduces the hyperproliferative phenotype of cystic cholangiocytes from an animal model of ARPKD

    Journal: Gastroenterology

    doi: 10.1053/j.gastro.2010.04.010

    Effect of Trpv4 activation on cystogenesis A, Representative images of cystic structures formed by PCK and normal bile ducts in the absence (Control) or presence of different Trpv4 activators: 5 μM 4αPDD, 0.5 μM 5’,6’-EET (EET) and 1 μM Nifedipine + 1μM AA (Nif+AA). Original magnification 40X B, Quantitative assessment of circumferential areas of cystic structures after 3 days of incubation showed that Trpv4 activation impaired PCK cyst expansion. Data are expressed as fold-increase compared to day 0. *p
    Figure Legend Snippet: Effect of Trpv4 activation on cystogenesis A, Representative images of cystic structures formed by PCK and normal bile ducts in the absence (Control) or presence of different Trpv4 activators: 5 μM 4αPDD, 0.5 μM 5’,6’-EET (EET) and 1 μM Nifedipine + 1μM AA (Nif+AA). Original magnification 40X B, Quantitative assessment of circumferential areas of cystic structures after 3 days of incubation showed that Trpv4 activation impaired PCK cyst expansion. Data are expressed as fold-increase compared to day 0. *p

    Techniques Used: Activation Assay, Incubation

    Effect of Trpv4 activation on cyst progression in vivo A, representative liver and kidney sections stained with picrosirius red from PCK rats treated with vehicle (n=6) or GSK1016790A (n=4). Bar, 2500 μm B, C quantification analysis of cystic and fibrotic area expressed as percentage of total parenchyma area. *p
    Figure Legend Snippet: Effect of Trpv4 activation on cyst progression in vivo A, representative liver and kidney sections stained with picrosirius red from PCK rats treated with vehicle (n=6) or GSK1016790A (n=4). Bar, 2500 μm B, C quantification analysis of cystic and fibrotic area expressed as percentage of total parenchyma area. *p

    Techniques Used: Activation Assay, In Vivo, Staining

    Trpv4 activation is associated with Akt activation and inhibition of active Erk and B-Raf NRC and PCK-CCL cells were treated with 4αPDD or GSK1016790A (GSK) and the activities of Akt, B-Raf and Erk were assessed. A, representative western blots of phospho- and total Akt isoforms and densitometric analysis showed that Trpv4 activation induced Akt activity in the PCK cells. B, PCK-CCL B-Raf activity, expressed as percentage of decrease compare to control. C, representative blots for phospho- and total-Erk isoforms and densitometric analysis suggested that Trpv4 activation induced a decrease in Erk activity in the PCK cells. Data represent percentage of ratio compared to each control (n=4). *p
    Figure Legend Snippet: Trpv4 activation is associated with Akt activation and inhibition of active Erk and B-Raf NRC and PCK-CCL cells were treated with 4αPDD or GSK1016790A (GSK) and the activities of Akt, B-Raf and Erk were assessed. A, representative western blots of phospho- and total Akt isoforms and densitometric analysis showed that Trpv4 activation induced Akt activity in the PCK cells. B, PCK-CCL B-Raf activity, expressed as percentage of decrease compare to control. C, representative blots for phospho- and total-Erk isoforms and densitometric analysis suggested that Trpv4 activation induced a decrease in Erk activity in the PCK cells. Data represent percentage of ratio compared to each control (n=4). *p

    Techniques Used: Activation Assay, Inhibition, Western Blot, Activity Assay

    Trpv4 is overexpressed in cystic cholangiocytes: confocal immunofluorescence and immunogold electron microscopy A, Confocal immunofluorescence images showing expression of Trpv4 in normal and PCK rats and in normal and ARPKD and ADPKD human liver samples (L, lumen; Trpv4 in green; acetylated α-tubulin in red; DAPI nuclear staining in blue). Original magnification 1000X (bars, 10 μm). B and C, Immunogold electron microscopy confirmed Trpv4 overexpression and showed its localization on apical and basolateral domains. Intracellular Trpv4 is significantly increased in PCK rat livers, while in normal liver gold particles were mainly on the apical domain. Bars, 500 nm; *p
    Figure Legend Snippet: Trpv4 is overexpressed in cystic cholangiocytes: confocal immunofluorescence and immunogold electron microscopy A, Confocal immunofluorescence images showing expression of Trpv4 in normal and PCK rats and in normal and ARPKD and ADPKD human liver samples (L, lumen; Trpv4 in green; acetylated α-tubulin in red; DAPI nuclear staining in blue). Original magnification 1000X (bars, 10 μm). B and C, Immunogold electron microscopy confirmed Trpv4 overexpression and showed its localization on apical and basolateral domains. Intracellular Trpv4 is significantly increased in PCK rat livers, while in normal liver gold particles were mainly on the apical domain. Bars, 500 nm; *p

    Techniques Used: Immunofluorescence, Electron Microscopy, Expressing, Staining, Over Expression

    Trpv4 activation increases intracellular calcium levels in PCK cholangiocytes A, 4αPDD induced intracellular calcium increases in a dose dependent manner. B, the alternative Trpv4 activators 5’,6’-EET and nifedipine (Nif) + arachidonic acid (AA) also increased intracellular calcium levels. (*p
    Figure Legend Snippet: Trpv4 activation increases intracellular calcium levels in PCK cholangiocytes A, 4αPDD induced intracellular calcium increases in a dose dependent manner. B, the alternative Trpv4 activators 5’,6’-EET and nifedipine (Nif) + arachidonic acid (AA) also increased intracellular calcium levels. (*p

    Techniques Used: Activation Assay

    Effect of Trpv4 activation on cholangiocyte proliferation A, Effect of Trpv4 activation by 4αPDD, the Nif+AA combination, 5’,6’-EET, and GSK1016790A on cell proliferation (n=10 for each activator) were analyzed by MTS assay over 3 days of culture. Data show the decrease in proliferation induced by the Trpv4 activators compared to control vehicle-treated cells. *p
    Figure Legend Snippet: Effect of Trpv4 activation on cholangiocyte proliferation A, Effect of Trpv4 activation by 4αPDD, the Nif+AA combination, 5’,6’-EET, and GSK1016790A on cell proliferation (n=10 for each activator) were analyzed by MTS assay over 3 days of culture. Data show the decrease in proliferation induced by the Trpv4 activators compared to control vehicle-treated cells. *p

    Techniques Used: Activation Assay, MTS Assay

    4αPDD inhibits cyst growth in 3D-culture in a Trpv4-dependent manner A, western blot of PCK cholangiocytes treated with scrambled (scr) or Trpv4-siRNA showed 80% decrease of Trpv4 protein induced by specific siRNA treatment. B, Cystic structures formed by PCK bile ducts where treated with a specific Trpv4-siRNA and then incubated in the absence or presence of 5 μM 4αPDD. Scrambled siRNA (scr) was used as a control. Quantitative assessment of circumferential areas of cystic structures after 3 days of incubation showed that Trpv4 is essential for the inhibition of cystogenesis induced by 4αPDD. Data are expressed as fold-increase compared to day 0. *p
    Figure Legend Snippet: 4αPDD inhibits cyst growth in 3D-culture in a Trpv4-dependent manner A, western blot of PCK cholangiocytes treated with scrambled (scr) or Trpv4-siRNA showed 80% decrease of Trpv4 protein induced by specific siRNA treatment. B, Cystic structures formed by PCK bile ducts where treated with a specific Trpv4-siRNA and then incubated in the absence or presence of 5 μM 4αPDD. Scrambled siRNA (scr) was used as a control. Quantitative assessment of circumferential areas of cystic structures after 3 days of incubation showed that Trpv4 is essential for the inhibition of cystogenesis induced by 4αPDD. Data are expressed as fold-increase compared to day 0. *p

    Techniques Used: Western Blot, Incubation, Inhibition

    Trpv4 is overexpressed in PCK cholangiocytes: qPCR and western blot A, Quantitative RT-PCR for Trpv4 on primary cultured cholangiocytes from normal and PCK rats (n=5). B, Representative western blot showing overexpression of Trpv4 in freshly isolated bile ducts from normal and PCK rats (n=5) and in cultured NRC and PCK-CCL (n=3). Data are shown as percentage of actin-normalized Trpv4 band compared to normal. *p
    Figure Legend Snippet: Trpv4 is overexpressed in PCK cholangiocytes: qPCR and western blot A, Quantitative RT-PCR for Trpv4 on primary cultured cholangiocytes from normal and PCK rats (n=5). B, Representative western blot showing overexpression of Trpv4 in freshly isolated bile ducts from normal and PCK rats (n=5) and in cultured NRC and PCK-CCL (n=3). Data are shown as percentage of actin-normalized Trpv4 band compared to normal. *p

    Techniques Used: Real-time Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR, Cell Culture, Over Expression, Isolation

    26) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    27) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    28) Product Images from "Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)"

    Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2017.00042

    PAR2 and TRPV4 expression in the hippocampus. Immunohistochemistry discloses the expression of PAR2 and TRPV4 in the hippocampus. A comparable expression pattern is observed: high levels of PAR2 and TRPV4 are detected in CA1 stratum pyramidale (pcl, pyramidal cell layer; oriens, stratum oriens; rad, stratum radiatum; la-mol, stratum lacunosum-moleculare). No pronounced colocalization between PAR2 and GFAP was detected. Scale bars: 100 and 10 μm, n = 9 slices out of three animals.
    Figure Legend Snippet: PAR2 and TRPV4 expression in the hippocampus. Immunohistochemistry discloses the expression of PAR2 and TRPV4 in the hippocampus. A comparable expression pattern is observed: high levels of PAR2 and TRPV4 are detected in CA1 stratum pyramidale (pcl, pyramidal cell layer; oriens, stratum oriens; rad, stratum radiatum; la-mol, stratum lacunosum-moleculare). No pronounced colocalization between PAR2 and GFAP was detected. Scale bars: 100 and 10 μm, n = 9 slices out of three animals.

    Techniques Used: Expressing, Immunohistochemistry

    PAR2 induces LTD through the activation of TRPV4. (A) Application of TRPV4-agonist (2 μM RN1747) causes LTD. (B) Removal of the TRPV4-agonist (2 μM RN1747) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of the TRPV4-antagonist (10 μM RN1734) the TRPV4-agonist is not able to induce synaptic depression. (D) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and TRPV4-agonist application induce similar levels of LTD. (E) LFS-induced LTD is not blocked by the TRPV4-antagonist. (F) Application of PAR2-agonist (10 μM AC55541) in presence of a TRPV4-antagonist (10 μM RN1734) blocks PAR2-induced LTD. (G) Application of TRPV4-agonist (2 μM RN1747) in presence of PAR2-antagonist (50 μM FSLLRY-NH 2 ) does not affect TRPV4-induced LTD. (H) Once PAR2-agonist mediated LTD is established, the TRPV4-agonist (2 μM RN1747) does not further de-potentiate a second pathway at adjusted response level (upward arrow). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiments, refer to text for statistics.
    Figure Legend Snippet: PAR2 induces LTD through the activation of TRPV4. (A) Application of TRPV4-agonist (2 μM RN1747) causes LTD. (B) Removal of the TRPV4-agonist (2 μM RN1747) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of the TRPV4-antagonist (10 μM RN1734) the TRPV4-agonist is not able to induce synaptic depression. (D) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and TRPV4-agonist application induce similar levels of LTD. (E) LFS-induced LTD is not blocked by the TRPV4-antagonist. (F) Application of PAR2-agonist (10 μM AC55541) in presence of a TRPV4-antagonist (10 μM RN1734) blocks PAR2-induced LTD. (G) Application of TRPV4-agonist (2 μM RN1747) in presence of PAR2-antagonist (50 μM FSLLRY-NH 2 ) does not affect TRPV4-induced LTD. (H) Once PAR2-agonist mediated LTD is established, the TRPV4-agonist (2 μM RN1747) does not further de-potentiate a second pathway at adjusted response level (upward arrow). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiments, refer to text for statistics.

    Techniques Used: Activation Assay

    TRPV4-mediated LTD depends on NMDAR-activity. (A) Similar to PAR2-induced LTD (c.f., Figures 1G,H ), the NMDAR-antagonist (50 μM APV) blocks TRPV4 (2 μM RN1747)-induced LTD, while (B) application of a TRPV4-agonist (2 μM RN1747) induces LTD in presence of the mGluR-antagonist (200 μM MCGP). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section.
    Figure Legend Snippet: TRPV4-mediated LTD depends on NMDAR-activity. (A) Similar to PAR2-induced LTD (c.f., Figures 1G,H ), the NMDAR-antagonist (50 μM APV) blocks TRPV4 (2 μM RN1747)-induced LTD, while (B) application of a TRPV4-agonist (2 μM RN1747) induces LTD in presence of the mGluR-antagonist (200 μM MCGP). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section.

    Techniques Used: Activity Assay

    29) Product Images from "Auricular Electroacupuncture Reduced Inflammation-Related Epilepsy Accompanied by Altered TRPA1, pPKCα, pPKCε, and pERk1/2 Signaling Pathways in Kainic Acid-Treated Rats"

    Article Title: Auricular Electroacupuncture Reduced Inflammation-Related Epilepsy Accompanied by Altered TRPA1, pPKCα, pPKCε, and pERk1/2 Signaling Pathways in Kainic Acid-Treated Rats

    Journal: Mediators of Inflammation

    doi: 10.1155/2014/493480

    TRPA1 and TRPV4 protein levels. Hippocampus lysates were immunoreacted with specific TRPA1 (a) and TRPV4 (b) antibodies. TRPA1, but not TRPV4, increased with KA injection as compared with the PBS group. TRPA1 protein levels were attenuated by electroacupuncture (EA) at EAR (auricular) as compared with the KA-induced groups. Serious results were not observed in ST36 (ST36-ST37) and sham groups. All statistic results were analyzed and plotted as bar chart in (c) and (d).
    Figure Legend Snippet: TRPA1 and TRPV4 protein levels. Hippocampus lysates were immunoreacted with specific TRPA1 (a) and TRPV4 (b) antibodies. TRPA1, but not TRPV4, increased with KA injection as compared with the PBS group. TRPA1 protein levels were attenuated by electroacupuncture (EA) at EAR (auricular) as compared with the KA-induced groups. Serious results were not observed in ST36 (ST36-ST37) and sham groups. All statistic results were analyzed and plotted as bar chart in (c) and (d).

    Techniques Used: Injection

    30) Product Images from "Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium"

    Article Title: Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium

    Journal: Acta Oto-Laryngologica

    doi: 10.3109/00016489.2010.489573

    The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p
    Figure Legend Snippet: The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p

    Techniques Used: Fluorescence

    Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.
    Figure Legend Snippet: Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.

    Techniques Used: Immunofluorescence, Labeling, Staining

    At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).
    Figure Legend Snippet: At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).

    Techniques Used: Immunofluorescence, Fluorescence, Staining

    31) Product Images from "Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium"

    Article Title: Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium

    Journal: Acta Oto-Laryngologica

    doi: 10.3109/00016489.2010.489573

    The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p
    Figure Legend Snippet: The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p

    Techniques Used: Fluorescence

    Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.
    Figure Legend Snippet: Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.

    Techniques Used: Immunofluorescence, Labeling, Staining

    At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).
    Figure Legend Snippet: At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).

    Techniques Used: Immunofluorescence, Fluorescence, Staining

    32) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    33) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    34) Product Images from "Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism"

    Article Title: Impairment in Function and Expression of TRPV4 in Dahl Salt-Sensitive Rats: Significance and Mechanism

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.147710

    Expression and regulation of TRPV4 in the kidney, DRG, and MA
    Figure Legend Snippet: Expression and regulation of TRPV4 in the kidney, DRG, and MA

    Techniques Used: Expressing

    Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without
    Figure Legend Snippet: Responses of MAP to bolus administration of 4α-PDD (2.5 mg/kg, iv) with or without TRPV4 blockade (RuR, 3 mg/kg, iv) in conscious DR or DS rats. A∼D, time course responses of MAP to bolus administration of 4α-PDD with or without

    Techniques Used:

    Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in DRG and MA of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P
    Figure Legend Snippet: Western blot analysis of protein expression of TRPV4 in the renal cortex and medulla of DR or DS rats. Values are mean ± SE (n=4∼5). *P

    Techniques Used: Western Blot, Expressing

    Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm
    Figure Legend Snippet: Confocal microscopic images of Cy3-labeled TRPV4 staining of mesenteric arteries in DR or DS rats fed a LS or HS diet. Negative controls were performed by omission of primary antibodies using DR rats fed a HS diet. Scale bars, 20μm

    Techniques Used: Labeling, Staining

    Effects of TRPV1 activation in the presence or absence of TRPV4 blockade
    Figure Legend Snippet: Effects of TRPV1 activation in the presence or absence of TRPV4 blockade

    Techniques Used: Activation Assay

    35) Product Images from "Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication"

    Article Title: Salt Intake Augments Hypotensive Effects of TRPV4: Functional Significance and Implication

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.108.117499

    Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Plasma calcitonin gene-related peptide (CGRP) and substance P (SP) levels 6 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P
    Figure Legend Snippet: MAP responses to intravenous injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without TRPV4 shRNA treatment in consious rats fed a NS or HS diet. Values are mean ± SE (n=4–6). * P

    Techniques Used: Injection, shRNA

    Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Polymerase chain reaction (PCR) analysis showing mRNA expressions of TRPV4 and TRPV1 in the renal medulla in HS- and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Polymerase Chain Reaction, shRNA

    Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P
    Figure Legend Snippet: Changes in mean arterial pressure (MAP) 5~7 min after bolus injection of 4α-PDD (2.5 mg/kg, iv, a TRPV4 activator) with or without SB 366791 (2 mg/kg, ip, a selective TRPV1 blocker), capsazepine (CAPZ, 3 mg/kg, iv, a selective TRPV1 blocker) and ruthenium red (RuR, 1 mg/kg and 3 mg/kg, iv, a TRPV4 blocker) in conscious rats fed a NS or HS diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–7). * P

    Techniques Used: Injection

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS and NS rats. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing

    Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P
    Figure Legend Snippet: Western blot analysis showing the TRPV4 protein expression in dorsal root ganglia (DRG), mesenteric resistant arteries (MA), the renal cortex and medulla in HS-and NS-treated rats with or without TRPV4 shRNA treatments. Values are mean ± SE (n=4–5). * P

    Techniques Used: Western Blot, Expressing, shRNA

    Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P
    Figure Legend Snippet: Time-course responses of mean arterial pressure (MAP, panel A) and heart rate (panel B) to bolus injection of 4α-PDD (1 mg/kg, 2.5 mg/kg, or 5 mg/kg, iv, a TRPV4 activator) in conscious rats fed a normal sodium diet. Vehicle: ethanol (5% v/v), Tween-80 (5% v/v) and saline, iv. Values are mean ± SE (n=5–8). * P

    Techniques Used: Injection

    36) Product Images from "Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension"

    Article Title: Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

    Journal: Journal of Lipid Research

    doi: 10.1016/j.jlr.2021.100145

    Most membrane TRPV4 does not partition into raft domains or interact with caveolar proteins. A: Western blot; detergent-free lipid raft isolation in primary TM cells. The supernatant fraction contains cytosolic proteins; fractions 1 and 2 (5% sucrose) contain nonlipid-raft membrane proteins, fractions 3–6 (5–35% sucrose) contain lipid raft membranes, and fractions 7 and 8 (pellet, 45% sucrose) contain unsuspended proteins and cell nuclei. TRPV4 protein is predominantly confined to fraction 2; fraction 4 is associated with flotillin-1 (48 kDa) and Cav-1 (22 kDa), whereas the supernatant/cytosol fraction associates with α-SMA (42 kDa). B: Coimmunoprecipitation. TRPV4-Cav-1 interaction assessed with the Cav-1 antibody for TRPV4 pulldown in control, MβCD, and MβCD:cholesterol-treated samples. Input bands, whole-cell lysate; bound bands, Cav-1-bound protein fraction; unbound bands, flow-through fractions. Cav-1-bound fractions show modest precipitation of the ∼75 kDa TRPV4 isoform. The absence of Cav-1 expression in unbound fractions confirms the quantitative immunoprecipitation of Cav-1. C, D: Immunohistochemistry, for control, MβCD, and MβCD:cholesterol-treated cells. C: Double immunolabeling for TRPV4 and Cav-1. TRPV4-ir (red arrowheads) does not colocalize with Cav1-ir puncta (green). D: Double immunolabeling for TRPV4 and flotillin. TRPV4-ir (red puncta) does not colocalize with flotillin-ir puncta (green). The inset is shown at higher magnification insets as supplemental Fig. S3 . The scale bar represents 20 μm.
    Figure Legend Snippet: Most membrane TRPV4 does not partition into raft domains or interact with caveolar proteins. A: Western blot; detergent-free lipid raft isolation in primary TM cells. The supernatant fraction contains cytosolic proteins; fractions 1 and 2 (5% sucrose) contain nonlipid-raft membrane proteins, fractions 3–6 (5–35% sucrose) contain lipid raft membranes, and fractions 7 and 8 (pellet, 45% sucrose) contain unsuspended proteins and cell nuclei. TRPV4 protein is predominantly confined to fraction 2; fraction 4 is associated with flotillin-1 (48 kDa) and Cav-1 (22 kDa), whereas the supernatant/cytosol fraction associates with α-SMA (42 kDa). B: Coimmunoprecipitation. TRPV4-Cav-1 interaction assessed with the Cav-1 antibody for TRPV4 pulldown in control, MβCD, and MβCD:cholesterol-treated samples. Input bands, whole-cell lysate; bound bands, Cav-1-bound protein fraction; unbound bands, flow-through fractions. Cav-1-bound fractions show modest precipitation of the ∼75 kDa TRPV4 isoform. The absence of Cav-1 expression in unbound fractions confirms the quantitative immunoprecipitation of Cav-1. C, D: Immunohistochemistry, for control, MβCD, and MβCD:cholesterol-treated cells. C: Double immunolabeling for TRPV4 and Cav-1. TRPV4-ir (red arrowheads) does not colocalize with Cav1-ir puncta (green). D: Double immunolabeling for TRPV4 and flotillin. TRPV4-ir (red puncta) does not colocalize with flotillin-ir puncta (green). The inset is shown at higher magnification insets as supplemental Fig. S3 . The scale bar represents 20 μm.

    Techniques Used: Western Blot, Isolation, Expressing, Immunoprecipitation, Immunohistochemistry, Immunolabeling

    Cholesterol depletion enhances TRPV4-mediated membrane currents in Xenopus oocytes. A: Representative current traces from TRPV4-expressing oocytes and uninjected control oocytes in control solution or after 45 min exposure to 50 μM MβCD. B: I/V curves of TRPV4-expressing oocytes exposed to control solution or MβCD, with uninjected oocytes in inset. Summarized currents obtained at −85 mV are shown in the lower inset. The magnitude of TRPV4-mediated currents (at V m = −85 mV) was compared using Student's t -test. ∗∗ P
    Figure Legend Snippet: Cholesterol depletion enhances TRPV4-mediated membrane currents in Xenopus oocytes. A: Representative current traces from TRPV4-expressing oocytes and uninjected control oocytes in control solution or after 45 min exposure to 50 μM MβCD. B: I/V curves of TRPV4-expressing oocytes exposed to control solution or MβCD, with uninjected oocytes in inset. Summarized currents obtained at −85 mV are shown in the lower inset. The magnitude of TRPV4-mediated currents (at V m = −85 mV) was compared using Student's t -test. ∗∗ P

    Techniques Used: Expressing

    Cholesterol depletion increases the number of TRPV4-ir puncta. TRPV4 immunolabeling of primary TM cells. Representative examples of (A) control, (B) 1 h treatment with MβCD, and (C) 1 h treatment with MβCD:cholesterol. Inset: zoomed-in region with TRPV4-ir puncta (arrow). D: Summary of three independent experiments, normalized for control cells. The number of TRPV4 puncta is upregulated after incubation with MβCD. ∗ P
    Figure Legend Snippet: Cholesterol depletion increases the number of TRPV4-ir puncta. TRPV4 immunolabeling of primary TM cells. Representative examples of (A) control, (B) 1 h treatment with MβCD, and (C) 1 h treatment with MβCD:cholesterol. Inset: zoomed-in region with TRPV4-ir puncta (arrow). D: Summary of three independent experiments, normalized for control cells. The number of TRPV4 puncta is upregulated after incubation with MβCD. ∗ P

    Techniques Used: Immunolabeling, Incubation

    Cholesterol depletion increases the amplitude of TRPV4 agonist-induced Ca 2+ signals. A, B: Ratiometric signals in Fura-2 AM-loaded cells. A: GSK101-induced elevations are increased in MβCD-treated cells (n = 8–10). B: Averaged data. MβCD (blue bar) augmented, whereas MβCD:cholesterol (green bar) reduced the amplitude of agonist-induced fluorescence. C: Fluorimetry, cell populations in 96 wells. About 5 nM GSK101 increased Fluo-4 fluorescence. Its effect was facilitated (∼12%) by MβCD (N = 4). ∗∗ P
    Figure Legend Snippet: Cholesterol depletion increases the amplitude of TRPV4 agonist-induced Ca 2+ signals. A, B: Ratiometric signals in Fura-2 AM-loaded cells. A: GSK101-induced elevations are increased in MβCD-treated cells (n = 8–10). B: Averaged data. MβCD (blue bar) augmented, whereas MβCD:cholesterol (green bar) reduced the amplitude of agonist-induced fluorescence. C: Fluorimetry, cell populations in 96 wells. About 5 nM GSK101 increased Fluo-4 fluorescence. Its effect was facilitated (∼12%) by MβCD (N = 4). ∗∗ P

    Techniques Used: Fluorescence

    37) Product Images from "Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium"

    Article Title: Expression of transient receptor potential channel vanilloid (TRPV) 1-4, melastin (TRPM) 5 and 8, and ankyrin (TRPA1) in the normal and methimazole-treated mouse olfactory epithelium

    Journal: Acta Oto-Laryngologica

    doi: 10.3109/00016489.2010.489573

    The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p
    Figure Legend Snippet: The fluorescence intensity to TRPV1 decreases significantly after 7 days but has normalized after 3 months. There is a significant difference in fluorescence intensity between 7 days and 1 month after. The intensity decreases significantly after 7 days and 1 month, with a significant difference between these two time points. The fluorescence intensities to TRPA1, TRPM5, TRPV3, and TRPV4 are significantly reduced after 1 month and 3 months, but have normalized after 3 months. The fluorescence intensity to TRPM8 is weaker but not significant after 7 days and 1 month. ** p

    Techniques Used: Fluorescence

    Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.
    Figure Legend Snippet: Immunofluorescent reaction to TRPV1 is evident in olfactory receptor neurons (ORNs). Supporting cells and basal cells are less intensely immunofluorescent. In lamina propria, nerve fibers (NFs) are immunofluorescent (TRPV1). Immunofluorescence to TRPV2 is evident in ORNs. Basal cells are markedly immunofluorescent. NFs are intensely immunofluorescent (TRPV2). Immunofluorescence to TRPV3 is observed in the ORNs. NFs show weak immunofluorescence (TRPV3). Immunofluorescence to TRPV4 is observed in the ORNs. In the lamina propria, TRPV4 labeling is weak and absent in NFs (TRPV4). The ORNs show immunofluorescence to TRPA1. Bowman's glands show marked immunofluorescence (TRPA1). Moderate immunofluorescence to TRPM5 is noted in the ORNs. Olfactory cilia are intensely immunofluorescent (TRPM5). Intense immunofluorescence to TRPM8 is observed in the ORNs. Olfactory cilia and basal cells are moderately immunofluorescent. NFs show marked immunofluorescence (TRPM8). Control staining without primary antibodies does not elicit immunofluorescence. Nuclei are counterstained with DAPI (control). O, ORNs; asterisk, nerve fiber; arrow, Bowman's gland; arrowhead, olfactory cilia.

    Techniques Used: Immunofluorescence, Labeling, Staining

    At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).
    Figure Legend Snippet: At 1 day after methimazole treatment (1d), there is a thin and disorganized neuroepithelium in the whole olfactory epithelium (OE; O). ORNs have almost completely disappeared. At 7 days later, OE shows signs of recovery but is still thin and disorganized (7d). At 1 month after treatment (1m), the OE is thicker, and after 3 months (3m) has regained its normal appearance and thickness. In general, the immunoreactivity of ORNs to TRPV1 is markedly reduced 7 days after methimazole treatment. A few cells show immunofluorescence (arrow). Immunofluorescence of ORNs has almost recovered (arrow) after 1 month and shows complete recovery after 3 months (TRPV1). Immunoreactivity of ORNs to TRPV2 is reduced in most ORNs, while a few cells show marked fluorescence (arrow) 7 days after the treatment. After 1 month, some cells show intense fluorescence (arrow), while others still show weaker fluorescence (TRPV2). Immunoreactivity of ORNs to TRPV3, TRPV4, and TRPA1 is reduced 7 days and 1 month after treatment. Immunoreactivity to TRPM5 is reduced after 7 days and 1 month. There is no fluorescence in olfactory cilia (arrow), while after 3 months, ORNs show normal fluorescence and there is intense staining in the olfactory cilia (arrow) (TRPM5). The immunoreactivity to TRPM8 is slightly weaker, but still strong after 7 days and 1 month, recovering to normal after 3 months (TRPM8).

    Techniques Used: Immunofluorescence, Fluorescence, Staining

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  • 93
    Alomone Labs antibody anti trpv4 extracellular
    PAR2 and <t>TRPV4</t> expression in the hippocampus. Immunohistochemistry discloses the expression of PAR2 and TRPV4 in the hippocampus. A comparable expression pattern is observed: high levels of PAR2 and TRPV4 are detected in CA1 stratum pyramidale (pcl, pyramidal cell layer; oriens, stratum oriens; rad, stratum radiatum; la-mol, stratum lacunosum-moleculare). No pronounced colocalization between PAR2 and GFAP was detected. Scale bars: 100 and 10 μm, n = 9 slices out of three animals.
    Antibody Anti Trpv4 Extracellular, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti rat trpv4
    Confocal microscope images of double-immunofluorescence staining of mesenteric resistant arteries (A, B, C, G, H, I) and DRG neurons (D, E, F). A, D and G: FITC-labeled <t>TRPV4</t> receptor staining (green). B and E: Cy3-labeled CGRP staining (red). C and F: colocalization of TRPV4 and CGRP (yellow). H: Cy3-labeled MaxiК channels staining (red). I: colocalization of TRPV4 and MaxiК channels (yellow). Blue arrows: the colocalized areas. Negative control not shown. Scale bars, 20 μm.
    Rabbit Anti Rat Trpv4, 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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    Alomone Labs trpv4
    FD20 content in serum. In Ctrl and DU groups, after treatment with <t>TRPV4</t> agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P
    Trpv4, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti trpv4 antibody
    Acute activation of the PKA signaling cascade promotes apical <t>TRPV4</t> translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that
    Anti Trpv4 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PAR2 and TRPV4 expression in the hippocampus. Immunohistochemistry discloses the expression of PAR2 and TRPV4 in the hippocampus. A comparable expression pattern is observed: high levels of PAR2 and TRPV4 are detected in CA1 stratum pyramidale (pcl, pyramidal cell layer; oriens, stratum oriens; rad, stratum radiatum; la-mol, stratum lacunosum-moleculare). No pronounced colocalization between PAR2 and GFAP was detected. Scale bars: 100 and 10 μm, n = 9 slices out of three animals.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

    doi: 10.3389/fnmol.2017.00042

    Figure Lengend Snippet: PAR2 and TRPV4 expression in the hippocampus. Immunohistochemistry discloses the expression of PAR2 and TRPV4 in the hippocampus. A comparable expression pattern is observed: high levels of PAR2 and TRPV4 are detected in CA1 stratum pyramidale (pcl, pyramidal cell layer; oriens, stratum oriens; rad, stratum radiatum; la-mol, stratum lacunosum-moleculare). No pronounced colocalization between PAR2 and GFAP was detected. Scale bars: 100 and 10 μm, n = 9 slices out of three animals.

    Article Snippet: Immunohistochemistry The following primary antibodies were used for immunodetection: goat anti-PAR2 (sc-8205, Santa Cruz, 1:25), rabbit anti-TRPV4 (ACC-124, Alomone Labs, 1:50), rabbit anti-PAR2 (APR-032, Alomone Labs 1:500) and mouse anti-GFAP (G3893, Sigma-Aldrich, 1:2000).

    Techniques: Expressing, Immunohistochemistry

    PAR2 induces LTD through the activation of TRPV4. (A) Application of TRPV4-agonist (2 μM RN1747) causes LTD. (B) Removal of the TRPV4-agonist (2 μM RN1747) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of the TRPV4-antagonist (10 μM RN1734) the TRPV4-agonist is not able to induce synaptic depression. (D) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and TRPV4-agonist application induce similar levels of LTD. (E) LFS-induced LTD is not blocked by the TRPV4-antagonist. (F) Application of PAR2-agonist (10 μM AC55541) in presence of a TRPV4-antagonist (10 μM RN1734) blocks PAR2-induced LTD. (G) Application of TRPV4-agonist (2 μM RN1747) in presence of PAR2-antagonist (50 μM FSLLRY-NH 2 ) does not affect TRPV4-induced LTD. (H) Once PAR2-agonist mediated LTD is established, the TRPV4-agonist (2 μM RN1747) does not further de-potentiate a second pathway at adjusted response level (upward arrow). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiments, refer to text for statistics.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

    doi: 10.3389/fnmol.2017.00042

    Figure Lengend Snippet: PAR2 induces LTD through the activation of TRPV4. (A) Application of TRPV4-agonist (2 μM RN1747) causes LTD. (B) Removal of the TRPV4-agonist (2 μM RN1747) following induction of LTD does not affect the stability of synaptic depression. (C) In presence of the TRPV4-antagonist (10 μM RN1734) the TRPV4-agonist is not able to induce synaptic depression. (D) In a two pathways experimental setting, low frequency stimulation (LFS, 1 Hz, 900 pulses) and TRPV4-agonist application induce similar levels of LTD. (E) LFS-induced LTD is not blocked by the TRPV4-antagonist. (F) Application of PAR2-agonist (10 μM AC55541) in presence of a TRPV4-antagonist (10 μM RN1734) blocks PAR2-induced LTD. (G) Application of TRPV4-agonist (2 μM RN1747) in presence of PAR2-antagonist (50 μM FSLLRY-NH 2 ) does not affect TRPV4-induced LTD. (H) Once PAR2-agonist mediated LTD is established, the TRPV4-agonist (2 μM RN1747) does not further de-potentiate a second pathway at adjusted response level (upward arrow). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section, n = 12 slices for each experiments, refer to text for statistics.

    Article Snippet: Immunohistochemistry The following primary antibodies were used for immunodetection: goat anti-PAR2 (sc-8205, Santa Cruz, 1:25), rabbit anti-TRPV4 (ACC-124, Alomone Labs, 1:50), rabbit anti-PAR2 (APR-032, Alomone Labs 1:500) and mouse anti-GFAP (G3893, Sigma-Aldrich, 1:2000).

    Techniques: Activation Assay

    TRPV4-mediated LTD depends on NMDAR-activity. (A) Similar to PAR2-induced LTD (c.f., Figures 1G,H ), the NMDAR-antagonist (50 μM APV) blocks TRPV4 (2 μM RN1747)-induced LTD, while (B) application of a TRPV4-agonist (2 μM RN1747) induces LTD in presence of the mGluR-antagonist (200 μM MCGP). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Protease Activated Receptor 2 (PAR2) Induces Long-Term Depression in the Hippocampus through Transient Receptor Potential Vanilloid 4 (TRPV4)

    doi: 10.3389/fnmol.2017.00042

    Figure Lengend Snippet: TRPV4-mediated LTD depends on NMDAR-activity. (A) Similar to PAR2-induced LTD (c.f., Figures 1G,H ), the NMDAR-antagonist (50 μM APV) blocks TRPV4 (2 μM RN1747)-induced LTD, while (B) application of a TRPV4-agonist (2 μM RN1747) induces LTD in presence of the mGluR-antagonist (200 μM MCGP). Averaged EPSP are plotted versus time. Representative traces at indicated times (a, b) are shown on top of each section.

    Article Snippet: Immunohistochemistry The following primary antibodies were used for immunodetection: goat anti-PAR2 (sc-8205, Santa Cruz, 1:25), rabbit anti-TRPV4 (ACC-124, Alomone Labs, 1:50), rabbit anti-PAR2 (APR-032, Alomone Labs 1:500) and mouse anti-GFAP (G3893, Sigma-Aldrich, 1:2000).

    Techniques: Activity Assay

    Confocal microscope images of double-immunofluorescence staining of mesenteric resistant arteries (A, B, C, G, H, I) and DRG neurons (D, E, F). A, D and G: FITC-labeled TRPV4 receptor staining (green). B and E: Cy3-labeled CGRP staining (red). C and F: colocalization of TRPV4 and CGRP (yellow). H: Cy3-labeled MaxiК channels staining (red). I: colocalization of TRPV4 and MaxiК channels (yellow). Blue arrows: the colocalized areas. Negative control not shown. Scale bars, 20 μm.

    Journal: Journal of hypertension

    Article Title: Hypotension Induced by TRPV4 Activation: Role of Ca2+-activated K+ Channels and Sensory Nerves

    doi: 10.1097/HJH.0b013e328332b865

    Figure Lengend Snippet: Confocal microscope images of double-immunofluorescence staining of mesenteric resistant arteries (A, B, C, G, H, I) and DRG neurons (D, E, F). A, D and G: FITC-labeled TRPV4 receptor staining (green). B and E: Cy3-labeled CGRP staining (red). C and F: colocalization of TRPV4 and CGRP (yellow). H: Cy3-labeled MaxiК channels staining (red). I: colocalization of TRPV4 and MaxiК channels (yellow). Blue arrows: the colocalized areas. Negative control not shown. Scale bars, 20 μm.

    Article Snippet: After washing with PBS, the arteries were incubated at 4 °C overnight in a primary antibody cocktail of goat anti-rat MaxiКβ (1:200, Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit anti-rat TRPV4 (1:100, Alomone labs, Jerusalem, Israel), followed by a secondary antibody cocktail of donkey anti-goat CY3 (1:136, Jackson ImmunoResearch labs, West Grove, PA) and donkey anti-rabbit FITC (1:136, Jackson ImmunoResearch labs, West Grove, PA) for 30 minutes.

    Techniques: Microscopy, Double Immunofluorescence Staining, Labeling, Staining, Negative Control

    FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Journal: BioMed Research International

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    doi: 10.1155/2022/2777882

    Figure Lengend Snippet: FD20 content in serum. In Ctrl and DU groups, after treatment with TRPV4 agonist GSK1016790 A or inhibitor HC067047, the content of FD20 in serum was detected. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Article Snippet: The major findings of this study are as follows: (1) the expression of TRPV4 was significantly increased in children with duodenal ulcer and had good diagnostic value in children with duodenal ulcer. (2) TRPV4 expression increased and calcium influx also increased in duodenal ulcer mice. (3) TRPV4 can promote intestinal epithelial permeability in mice with duodenal ulcer. (4) Intestinal permeability mediated by TRPV4 can affect the entry of cytokines into the blood and exacerbate the progression of ulcers.

    Techniques:

    The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Journal: BioMed Research International

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    doi: 10.1155/2022/2777882

    Figure Lengend Snippet: The content of inflammatory factor TNF- α in tissues and serum. (a) In the Ctrl and DU groups, serum TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. (b) In the Ctrl and DU groups, duodenal tissue TNF- α content changes after treatment with TRPV4 agonists GSK1016790 A or inhibitor HC067047. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Article Snippet: The major findings of this study are as follows: (1) the expression of TRPV4 was significantly increased in children with duodenal ulcer and had good diagnostic value in children with duodenal ulcer. (2) TRPV4 expression increased and calcium influx also increased in duodenal ulcer mice. (3) TRPV4 can promote intestinal epithelial permeability in mice with duodenal ulcer. (4) Intestinal permeability mediated by TRPV4 can affect the entry of cytokines into the blood and exacerbate the progression of ulcers.

    Techniques:

    TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P

    Journal: BioMed Research International

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    doi: 10.1155/2022/2777882

    Figure Lengend Snippet: TRPV4 is highly expressed in children with duodenal ulcer and has good diagnostic value. (a) and (b) Immunohistochemistry representative images (a) and summary (b) data showing the expression of TRPV4 in children with duodenal ulcer; Ctrl: nonduodenal ulcer; DU: duodenal ulcer. (c) ROC curve analyses of TRPV4. n = 11, ∗ P

    Article Snippet: The major findings of this study are as follows: (1) the expression of TRPV4 was significantly increased in children with duodenal ulcer and had good diagnostic value in children with duodenal ulcer. (2) TRPV4 expression increased and calcium influx also increased in duodenal ulcer mice. (3) TRPV4 can promote intestinal epithelial permeability in mice with duodenal ulcer. (4) Intestinal permeability mediated by TRPV4 can affect the entry of cytokines into the blood and exacerbate the progression of ulcers.

    Techniques: Diagnostic Assay, Immunohistochemistry, Expressing

    Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Journal: BioMed Research International

    Article Title: Role of TRPV4 in the Diagnosis and Treatment of Helicobacter pylori Infection in Children with Duodenal Ulcers

    doi: 10.1155/2022/2777882

    Figure Lengend Snippet: Increased TRPV4 expression and enhanced calcium influx in duodenal ulcer mice. (a) Representative images showing hematoxylin-eosin staining of duodenum tissue. (b) and (c) Representative images (b) and summary (c) data showing the expression of TRPV4 in duodenal ulcer mice. (d) and (e) Representative traces (d) and summarized data (e) showing the changes in the intracellular Ca 2+ concentration of duodenal tissue between DU and Ctrl group. Ctrl: nonduodenal ulcer; DU: duodenal ulcer. n = 3, ∗ P

    Article Snippet: The major findings of this study are as follows: (1) the expression of TRPV4 was significantly increased in children with duodenal ulcer and had good diagnostic value in children with duodenal ulcer. (2) TRPV4 expression increased and calcium influx also increased in duodenal ulcer mice. (3) TRPV4 can promote intestinal epithelial permeability in mice with duodenal ulcer. (4) Intestinal permeability mediated by TRPV4 can affect the entry of cytokines into the blood and exacerbate the progression of ulcers.

    Techniques: Expressing, Mouse Assay, Staining, Concentration Assay

    Acute activation of the PKA signaling cascade promotes apical TRPV4 translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Acute activation of the PKA signaling cascade promotes apical TRPV4 translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques: Activation Assay, Translocation Assay

    Regulation of mechanosensitive [Ca 2+ ] i responses by PKA and PKC cascades occurs in a TRPV4-dependent manner. A , average time course of changes in [Ca 2+ ] i levels in response to a 10-fold elevation in flow over the apical surface ( gray bars ) for individual

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Regulation of mechanosensitive [Ca 2+ ] i responses by PKA and PKC cascades occurs in a TRPV4-dependent manner. A , average time course of changes in [Ca 2+ ] i levels in response to a 10-fold elevation in flow over the apical surface ( gray bars ) for individual

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques: Flow Cytometry

    Distinct effects of PKC- and PKA-dependent signaling cascades on subcellular TRPV4 localization in distal nephron cells. Shown are representative confocal plane micrographs (axes are shown) and corresponding cross-sections (indicated by arrows ) demonstrating

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Distinct effects of PKC- and PKA-dependent signaling cascades on subcellular TRPV4 localization in distal nephron cells. Shown are representative confocal plane micrographs (axes are shown) and corresponding cross-sections (indicated by arrows ) demonstrating

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques: