anti trpv4 extracellular antibody  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93

    Structured Review

    Alomone Labs anti trpv4 extracellular antibody
    A) Immunofluorescent images of ATDC5 cells on normal gels (upper panel) treated with 30nM GSK101 for 28 hr and severe OA gels with control treatment (lower panel) stained for the nucleus (Hoechst, blue), f-actin (purple), and <t>TRPV4</t> (white). The left panel shows the composite images of all stains (scale bar =10 μm). The middle panel shows nuclear and f-actin staining, and the right panel shows TRPV4 and nuclear staining. B) TRPV4 fluorescence of ATDC5 cells on normal, OA, and severe OA gels during control (dark blue) and 28 hr of 30 nM GSK101 (light blue) treatments. f-actin was used to draw ROI around the cells to measure TRPV4 fluorescence from max projections of z-stack images. Error Bars are ± SEM, * indicates p-value
    Anti Trpv4 Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti trpv4 extracellular antibody/product/Alomone Labs
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    anti trpv4 extracellular antibody - by Bioz Stars, 2022-08
    93/100 stars

    Images

    1) Product Images from "Extracellular Matrix Stiffness Alters TRPV4 Regulation in Chondrocytes"

    Article Title: Extracellular Matrix Stiffness Alters TRPV4 Regulation in Chondrocytes

    Journal: bioRxiv

    doi: 10.1101/2021.09.14.460172

    A) Immunofluorescent images of ATDC5 cells on normal gels (upper panel) treated with 30nM GSK101 for 28 hr and severe OA gels with control treatment (lower panel) stained for the nucleus (Hoechst, blue), f-actin (purple), and TRPV4 (white). The left panel shows the composite images of all stains (scale bar =10 μm). The middle panel shows nuclear and f-actin staining, and the right panel shows TRPV4 and nuclear staining. B) TRPV4 fluorescence of ATDC5 cells on normal, OA, and severe OA gels during control (dark blue) and 28 hr of 30 nM GSK101 (light blue) treatments. f-actin was used to draw ROI around the cells to measure TRPV4 fluorescence from max projections of z-stack images. Error Bars are ± SEM, * indicates p-value
    Figure Legend Snippet: A) Immunofluorescent images of ATDC5 cells on normal gels (upper panel) treated with 30nM GSK101 for 28 hr and severe OA gels with control treatment (lower panel) stained for the nucleus (Hoechst, blue), f-actin (purple), and TRPV4 (white). The left panel shows the composite images of all stains (scale bar =10 μm). The middle panel shows nuclear and f-actin staining, and the right panel shows TRPV4 and nuclear staining. B) TRPV4 fluorescence of ATDC5 cells on normal, OA, and severe OA gels during control (dark blue) and 28 hr of 30 nM GSK101 (light blue) treatments. f-actin was used to draw ROI around the cells to measure TRPV4 fluorescence from max projections of z-stack images. Error Bars are ± SEM, * indicates p-value

    Techniques Used: Staining, Fluorescence

    A) mRNA expression of anabolic factors (Sox-9, aggrecan, col2 and TRPV4) in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. B) mRNA expression of catabolic chondrocyte factors, collagen I and MMP-13 in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. Fold change was normalized to rplp0 expression for each sample and then normalized to the normal control condition. C) Western blotting shows TRPV4 protein levels for ATDC5 cells on normal, OA, and severe OA gels during control treatment. The bar graph represents the normalized TRPV4 protein level to GAPDH for each sample. Error bars are ± SEM, * indicates significance with a p-value
    Figure Legend Snippet: A) mRNA expression of anabolic factors (Sox-9, aggrecan, col2 and TRPV4) in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. B) mRNA expression of catabolic chondrocyte factors, collagen I and MMP-13 in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. Fold change was normalized to rplp0 expression for each sample and then normalized to the normal control condition. C) Western blotting shows TRPV4 protein levels for ATDC5 cells on normal, OA, and severe OA gels during control treatment. The bar graph represents the normalized TRPV4 protein level to GAPDH for each sample. Error bars are ± SEM, * indicates significance with a p-value

    Techniques Used: Expressing, Western Blot

    2) Product Images from "Omega-3 Fatty Acids Modulate TRPV4 Function Through Plasma Membrane Remodeling"

    Article Title: Omega-3 Fatty Acids Modulate TRPV4 Function Through Plasma Membrane Remodeling

    Journal: Cell reports

    doi: 10.1016/j.celrep.2017.09.029

    GSK101 elicits withdrawal responses in rat TRPV4-expressing worms. (A) Schematic representation of the withdrawal responses after addition of GSK101 drop in front of freely moving worms. (B) GSK101 dose-response profile for wild-type (WT [N2]) and TRPV4-expressing worms. (C) Inhibition of GSK101-mediated withdrawal responses in TRPV4 worms by HC067047 (2 µM). (D) Withdrawal responses elicited by 4α-Phorbol in WT and TRPV4 worms. (E) Withdrawal responses elicited by 1 M glycerol and nose touch in WT, osm9 , and TRPV4; osm9 strains. Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p
    Figure Legend Snippet: GSK101 elicits withdrawal responses in rat TRPV4-expressing worms. (A) Schematic representation of the withdrawal responses after addition of GSK101 drop in front of freely moving worms. (B) GSK101 dose-response profile for wild-type (WT [N2]) and TRPV4-expressing worms. (C) Inhibition of GSK101-mediated withdrawal responses in TRPV4 worms by HC067047 (2 µM). (D) Withdrawal responses elicited by 4α-Phorbol in WT and TRPV4 worms. (E) Withdrawal responses elicited by 1 M glycerol and nose touch in WT, osm9 , and TRPV4; osm9 strains. Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p

    Techniques Used: Expressing, Inhibition

    EPA supplementation enhances TRPV4 activity in HMVEC. (A) Representative whole-cell patch-clamp recordings (+80 mV) of control and EPA (100 µM)-treated HMVEC challenged with GSK101 (100 nM) and HC067047 (10 µM). (B) Box plots show the mean, median, standard deviation, and standard error of the mean from TRPV4 currents (I GSK101 - I HC / pF) obtained by whole-cell patch-clamp recordings (+80 mV) of control, EPA-, and ω -6 AA-treated HMVEC. (C) Left, representative current-voltage relationships determined by whole-cell patch-clamp recording of control and EPA (100 µM)-treated HMVEC challenged with GSK101 (100 nM) in the presence of 5 mM Ca 2+ . Right, bar graph of peak currents (at +80 mV) relative to the currents after 5 min of exposure to GSK101 (I max /I 5 min ). Bars are mean ± SEM. (D) HMVEC were challenged with isosmotic (IB, 320 mOsm), hyposmotic (HB, 240 mOsm), and GSK101 (100 nM) solutions and analyzed for their responses using Ca 2+ imaging (Fluo-4 AM); color bar indicates relative change in fluorescence intensity. Control and EPA (100–300 µM)-treated HMVEC were analyzed from 5 independent preparations. (E) Representative traces corresponding to normalized (ΔF/F) intensity changes of individual cells shown in (D). (F) Area under the curve of control and EPA-treated HMVEC challenged with hyposmotic buffer. Bars are mean ± SEM. The number of endothelial cells measured is indicated below the boxes and inside the bars. The asterisks indicate values significantly different from control. *** p
    Figure Legend Snippet: EPA supplementation enhances TRPV4 activity in HMVEC. (A) Representative whole-cell patch-clamp recordings (+80 mV) of control and EPA (100 µM)-treated HMVEC challenged with GSK101 (100 nM) and HC067047 (10 µM). (B) Box plots show the mean, median, standard deviation, and standard error of the mean from TRPV4 currents (I GSK101 - I HC / pF) obtained by whole-cell patch-clamp recordings (+80 mV) of control, EPA-, and ω -6 AA-treated HMVEC. (C) Left, representative current-voltage relationships determined by whole-cell patch-clamp recording of control and EPA (100 µM)-treated HMVEC challenged with GSK101 (100 nM) in the presence of 5 mM Ca 2+ . Right, bar graph of peak currents (at +80 mV) relative to the currents after 5 min of exposure to GSK101 (I max /I 5 min ). Bars are mean ± SEM. (D) HMVEC were challenged with isosmotic (IB, 320 mOsm), hyposmotic (HB, 240 mOsm), and GSK101 (100 nM) solutions and analyzed for their responses using Ca 2+ imaging (Fluo-4 AM); color bar indicates relative change in fluorescence intensity. Control and EPA (100–300 µM)-treated HMVEC were analyzed from 5 independent preparations. (E) Representative traces corresponding to normalized (ΔF/F) intensity changes of individual cells shown in (D). (F) Area under the curve of control and EPA-treated HMVEC challenged with hyposmotic buffer. Bars are mean ± SEM. The number of endothelial cells measured is indicated below the boxes and inside the bars. The asterisks indicate values significantly different from control. *** p

    Techniques Used: Activity Assay, Patch Clamp, Standard Deviation, Imaging, Fluorescence

    EPA supplementation increases ω -3 fatty acid eicosanoid derivatives in HMVEC and does not affect TRPV4 expression and trafficking. (A) EPA and ω -6 AA content in control and EPA (100 µM)-treated HMVEC, as determined by LC-MS. (B) ω .
    Figure Legend Snippet: EPA supplementation increases ω -3 fatty acid eicosanoid derivatives in HMVEC and does not affect TRPV4 expression and trafficking. (A) EPA and ω -6 AA content in control and EPA (100 µM)-treated HMVEC, as determined by LC-MS. (B) ω .

    Techniques Used: Expressing, Liquid Chromatography with Mass Spectroscopy

    EPA and 17,18-EEQ fully restore TRPV4 function in C. elegans . (A) GSK101-mediated withdrawal responses of TRPV4 and TRPV4; fat 3 mutants after worms were fed with specified PUFAs and eicosanoid derivatives (200 µM). Dotted red and blue lines represent the 20% and 45% thresholds for positive and intermediate responses, respectively. (B) Schematic representation of the effect of ETYA (non-metabolizable analogue of ω -6 AA) in worms. (C) Top inset, ω -6 PUFAs present in fat-1 and fat-1fat-4 . Bottom, withdrawal responses elicited by GSK101 in TRPV4; fat-3 , TRPV4; fat-3 supplemented with ETYA, TRPV4; fat-1, TRPV4; fat-1fat-4, and TRPV4; fat-1fat-4 supplemented with ω -6 AA. (D) Withdrawal responses elicited by 1 M glycerol and nose touch in TRPV4; osm9fat-3 mutants after being fed with EPA (200 µM). Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p
    Figure Legend Snippet: EPA and 17,18-EEQ fully restore TRPV4 function in C. elegans . (A) GSK101-mediated withdrawal responses of TRPV4 and TRPV4; fat 3 mutants after worms were fed with specified PUFAs and eicosanoid derivatives (200 µM). Dotted red and blue lines represent the 20% and 45% thresholds for positive and intermediate responses, respectively. (B) Schematic representation of the effect of ETYA (non-metabolizable analogue of ω -6 AA) in worms. (C) Top inset, ω -6 PUFAs present in fat-1 and fat-1fat-4 . Bottom, withdrawal responses elicited by GSK101 in TRPV4; fat-3 , TRPV4; fat-3 supplemented with ETYA, TRPV4; fat-1, TRPV4; fat-1fat-4, and TRPV4; fat-1fat-4 supplemented with ω -6 AA. (D) Withdrawal responses elicited by 1 M glycerol and nose touch in TRPV4; osm9fat-3 mutants after being fed with EPA (200 µM). Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p

    Techniques Used:

    PUFAs are required for TRPV4 function in C. elegans ). LA, linolenic acid; γLA, γ-linolenic acid; DγLA, dihomo-linolenic acid; ω -6 AA, arachidonic acid; EET, epoxy-eicosatrienoic acid; ALA, α-linolenic acid; STA, stearidonic acid; ω -3 AA; EPA, eicosapentaenoic acid; EEQ, 17’18’-epoxy eicosatetraenoic acid. (B) Withdrawal responses elicited by GSK101 in WT, TRPV4, TRPV4; fat-3 , and TRPV4; fat-3 worms supplemented with PUFAs. (C) Withdrawal responses elicited by 4α-Phorbol in WT, TRPV4, and TRPV4; fat-3 worms. (D) Withdrawal responses elicited by GSK101 in TRPV4 and TRPV4; fat-4 worms. (E) Withdrawal responses elicited by 1 M glycerol and nose touch in TRPV4; osm9 and TRPV4; osm9fat-3 strains. (F) Representative micrographs of TRPV4::GFP and TRPV4::GFP; fat-3 ASH neurons. (G) Box plots show the mean, median, and the 75 th to 25 th percentiles of the fluorescence intensity analysis from images in (F). The number of neurons imaged during 2 sessions is indicated below the boxes. (H) Schematic representation of the phospholipid synthesis. (I) GSK101 withdrawal responses after knocking down the expression of mboa-6 in TRPV4 worms. Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p .
    Figure Legend Snippet: PUFAs are required for TRPV4 function in C. elegans ). LA, linolenic acid; γLA, γ-linolenic acid; DγLA, dihomo-linolenic acid; ω -6 AA, arachidonic acid; EET, epoxy-eicosatrienoic acid; ALA, α-linolenic acid; STA, stearidonic acid; ω -3 AA; EPA, eicosapentaenoic acid; EEQ, 17’18’-epoxy eicosatetraenoic acid. (B) Withdrawal responses elicited by GSK101 in WT, TRPV4, TRPV4; fat-3 , and TRPV4; fat-3 worms supplemented with PUFAs. (C) Withdrawal responses elicited by 4α-Phorbol in WT, TRPV4, and TRPV4; fat-3 worms. (D) Withdrawal responses elicited by GSK101 in TRPV4 and TRPV4; fat-4 worms. (E) Withdrawal responses elicited by 1 M glycerol and nose touch in TRPV4; osm9 and TRPV4; osm9fat-3 strains. (F) Representative micrographs of TRPV4::GFP and TRPV4::GFP; fat-3 ASH neurons. (G) Box plots show the mean, median, and the 75 th to 25 th percentiles of the fluorescence intensity analysis from images in (F). The number of neurons imaged during 2 sessions is indicated below the boxes. (H) Schematic representation of the phospholipid synthesis. (I) GSK101 withdrawal responses after knocking down the expression of mboa-6 in TRPV4 worms. Bars are mean ± SEM, the number of worms tested during 3 assays sessions is indicated inside the bars. The asterisks indicate values significantly different from control. *** p .

    Techniques Used: Fluorescence, Expressing

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 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
    https://www.bioz.com/result/antibody anti trpv4 extracellular/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibody anti trpv4 extracellular - by Bioz Stars, 2022-08
    93/100 stars
      Buy from Supplier

    95
    Alomone Labs anti mouse trpv4
    Transfection of large mouse cholangiocytes (MLC) with plasmid for transient receptor potential vanilloid member 4 <t>(TRPV4)</t> small interfering (si)RNA decreases protein and RNA expression. A : representative Western blot demonstrating change in TRPV4 protein levels in cells transfected with nontargeting siRNA (scramble) and cells transfected with TRPV4 siRNA. β-Actin was used as loading control. B : cumulative data demonstrating change in TRPV4 protein ( left ) and mRNA ( right ) levels in control cells, cells transfected with nontargeting siRNA (scramble), and cells transfected with TRPV4 siRNA (* P
    Anti Mouse 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
    https://www.bioz.com/result/anti mouse trpv4/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mouse trpv4 - by Bioz Stars, 2022-08
    95/100 stars
      Buy from Supplier

    Image Search Results


    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

    TRPV4 was downregulated in mature CD11c + BMDCs. ( A ) Concentration dependence of immature (black bars) and mature (dark cyan bars) CD11c + BMDC responding fraction. ***, p

    Journal: International Journal of Molecular Sciences

    Article Title: Expression and Functional Role of TRPV4 in Bone Marrow-Derived CD11c+ Cells

    doi: 10.3390/ijms20143378

    Figure Lengend Snippet: TRPV4 was downregulated in mature CD11c + BMDCs. ( A ) Concentration dependence of immature (black bars) and mature (dark cyan bars) CD11c + BMDC responding fraction. ***, p

    Article Snippet: Cells were blocked with antimouse CD16/32 polyclonal antibody (1 µg/mL, eBioscience) in 5% sheep serum (Sigma-Aldrich) for 3 h. After two rinsing steps with PBS, cells were incubated overnight at 4 °C with a rabbit anti-TRPV4 antibody (1:200, ACC-124, Alomone labs, Jerusalem, Israel).

    Techniques: Concentration Assay

    TRPV4-deficient BMDCs exhibited impaired FcR-dependent phagocytosis. ( A ) Representative confocal images of wild-type and Trpv4 KO BMDCs after treatment with uncoated or IgG-coated fluorescent microspheres. Scale bar, 20 µm. ( B ) Percentage of cells with internalized beads. Data were collected from 10 randomly selected fields per condition from three independent experiments. ***, p

    Journal: International Journal of Molecular Sciences

    Article Title: Expression and Functional Role of TRPV4 in Bone Marrow-Derived CD11c+ Cells

    doi: 10.3390/ijms20143378

    Figure Lengend Snippet: TRPV4-deficient BMDCs exhibited impaired FcR-dependent phagocytosis. ( A ) Representative confocal images of wild-type and Trpv4 KO BMDCs after treatment with uncoated or IgG-coated fluorescent microspheres. Scale bar, 20 µm. ( B ) Percentage of cells with internalized beads. Data were collected from 10 randomly selected fields per condition from three independent experiments. ***, p

    Article Snippet: Cells were blocked with antimouse CD16/32 polyclonal antibody (1 µg/mL, eBioscience) in 5% sheep serum (Sigma-Aldrich) for 3 h. After two rinsing steps with PBS, cells were incubated overnight at 4 °C with a rabbit anti-TRPV4 antibody (1:200, ACC-124, Alomone labs, Jerusalem, Israel).

    Techniques:

    TRPV4 was dispensable in the differentiation of CD11c + BMDCs. ( A ) Color-coded two-dimensional t-distributed stochastic neighbor embedding (tSNE) representations of the total bone marrow-derived cell population (20,000 cells) defined by the surface markers CD11b, CD11c, and F4/80. ( B ) Histograms showing surface expression of the indicated markers in bone marrow-derived cells from wild-type (WT, black traces) and Trpv4 knockout (KO, red traces) mice. The shaded histograms represent specificity (fluorescence minus one) controls. The bar graph shows the percentage of different cell populations present in total bone marrow-derived cell cultures defined by the surface expression of CD11b, CD11c, and F4/80. The data are represented as mean ± SEM of nine independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Expression and Functional Role of TRPV4 in Bone Marrow-Derived CD11c+ Cells

    doi: 10.3390/ijms20143378

    Figure Lengend Snippet: TRPV4 was dispensable in the differentiation of CD11c + BMDCs. ( A ) Color-coded two-dimensional t-distributed stochastic neighbor embedding (tSNE) representations of the total bone marrow-derived cell population (20,000 cells) defined by the surface markers CD11b, CD11c, and F4/80. ( B ) Histograms showing surface expression of the indicated markers in bone marrow-derived cells from wild-type (WT, black traces) and Trpv4 knockout (KO, red traces) mice. The shaded histograms represent specificity (fluorescence minus one) controls. The bar graph shows the percentage of different cell populations present in total bone marrow-derived cell cultures defined by the surface expression of CD11b, CD11c, and F4/80. The data are represented as mean ± SEM of nine independent experiments.

    Article Snippet: Cells were blocked with antimouse CD16/32 polyclonal antibody (1 µg/mL, eBioscience) in 5% sheep serum (Sigma-Aldrich) for 3 h. After two rinsing steps with PBS, cells were incubated overnight at 4 °C with a rabbit anti-TRPV4 antibody (1:200, ACC-124, Alomone labs, Jerusalem, Israel).

    Techniques: Derivative Assay, Expressing, Knock-Out, Mouse Assay, Fluorescence

    TRPV4 was functionally expressed in CD11c + bone marrow-derived cells (BMDCs). ( A ) Expression profile of selected Trp genes in the total granulocyte-macrophage colony-stimulating (GM-CSF)-differentiated bone marrow-derived cell population (black bars) and in CD11c + -purified BMDCs (light gray). Values are relative to GAPDH expression. ( B ) Confocal image of CD11c + BMDCs stained with an anti-TRPV4 antibody (red). The blue color corresponds to nuclear staining with DAPI. ( C–E ) Representative traces of intracellular Ca 2+ concentration in CD11c + BMDCs showing the effects of 300 nM of GSK1016790A (GSK). ATP (100 μM) was used as a positive control for intracellular Ca 2+ increase. The TRPV4 antagonist HC067047 was used at 10 μM. ( F ) Percentage of CD11c + BMDCs responding to the indicated stimulus. GSK, GSK1016790A (300 nM); HC, HC067047 (1 µM); Ca 2+ -free, Krebs with nominal [Ca 2+ ] supplemented with 2.5 mM EDTA; Caps, Capsaicin (1 nM); THC, trans-Δ 9 -tetrahydrocannabinol (10 µM). The responding fraction is indicated within each bar. ***, p

    Journal: International Journal of Molecular Sciences

    Article Title: Expression and Functional Role of TRPV4 in Bone Marrow-Derived CD11c+ Cells

    doi: 10.3390/ijms20143378

    Figure Lengend Snippet: TRPV4 was functionally expressed in CD11c + bone marrow-derived cells (BMDCs). ( A ) Expression profile of selected Trp genes in the total granulocyte-macrophage colony-stimulating (GM-CSF)-differentiated bone marrow-derived cell population (black bars) and in CD11c + -purified BMDCs (light gray). Values are relative to GAPDH expression. ( B ) Confocal image of CD11c + BMDCs stained with an anti-TRPV4 antibody (red). The blue color corresponds to nuclear staining with DAPI. ( C–E ) Representative traces of intracellular Ca 2+ concentration in CD11c + BMDCs showing the effects of 300 nM of GSK1016790A (GSK). ATP (100 μM) was used as a positive control for intracellular Ca 2+ increase. The TRPV4 antagonist HC067047 was used at 10 μM. ( F ) Percentage of CD11c + BMDCs responding to the indicated stimulus. GSK, GSK1016790A (300 nM); HC, HC067047 (1 µM); Ca 2+ -free, Krebs with nominal [Ca 2+ ] supplemented with 2.5 mM EDTA; Caps, Capsaicin (1 nM); THC, trans-Δ 9 -tetrahydrocannabinol (10 µM). The responding fraction is indicated within each bar. ***, p

    Article Snippet: Cells were blocked with antimouse CD16/32 polyclonal antibody (1 µg/mL, eBioscience) in 5% sheep serum (Sigma-Aldrich) for 3 h. After two rinsing steps with PBS, cells were incubated overnight at 4 °C with a rabbit anti-TRPV4 antibody (1:200, ACC-124, Alomone labs, Jerusalem, Israel).

    Techniques: Derivative Assay, Expressing, Purification, Staining, Concentration Assay, Positive Control

    LPS-induced cytokine production occurred independently of TRPV4. ( A ) Representative confocal immunofluorescence microscopy images of fixed BMDCs untreated or treated with LPS (100 ng/mL). Cell stainings correspond to NF-κB p65 (red) and DAPI (nuclear, blue). Scale bar, 10 µm. The average linear intensity along the gray rectangle is represented next to the corresponding image. ( B ) Percentage of the total nuclear area stained by NF-κB p65 staining. The horizontal bar represents the mean. ***, p

    Journal: International Journal of Molecular Sciences

    Article Title: Expression and Functional Role of TRPV4 in Bone Marrow-Derived CD11c+ Cells

    doi: 10.3390/ijms20143378

    Figure Lengend Snippet: LPS-induced cytokine production occurred independently of TRPV4. ( A ) Representative confocal immunofluorescence microscopy images of fixed BMDCs untreated or treated with LPS (100 ng/mL). Cell stainings correspond to NF-κB p65 (red) and DAPI (nuclear, blue). Scale bar, 10 µm. The average linear intensity along the gray rectangle is represented next to the corresponding image. ( B ) Percentage of the total nuclear area stained by NF-κB p65 staining. The horizontal bar represents the mean. ***, p

    Article Snippet: Cells were blocked with antimouse CD16/32 polyclonal antibody (1 µg/mL, eBioscience) in 5% sheep serum (Sigma-Aldrich) for 3 h. After two rinsing steps with PBS, cells were incubated overnight at 4 °C with a rabbit anti-TRPV4 antibody (1:200, ACC-124, Alomone labs, Jerusalem, Israel).

    Techniques: Immunofluorescence, Microscopy, Staining

    A) Immunofluorescent images of ATDC5 cells on normal gels (upper panel) treated with 30nM GSK101 for 28 hr and severe OA gels with control treatment (lower panel) stained for the nucleus (Hoechst, blue), f-actin (purple), and TRPV4 (white). The left panel shows the composite images of all stains (scale bar =10 μm). The middle panel shows nuclear and f-actin staining, and the right panel shows TRPV4 and nuclear staining. B) TRPV4 fluorescence of ATDC5 cells on normal, OA, and severe OA gels during control (dark blue) and 28 hr of 30 nM GSK101 (light blue) treatments. f-actin was used to draw ROI around the cells to measure TRPV4 fluorescence from max projections of z-stack images. Error Bars are ± SEM, * indicates p-value

    Journal: bioRxiv

    Article Title: Extracellular Matrix Stiffness Alters TRPV4 Regulation in Chondrocytes

    doi: 10.1101/2021.09.14.460172

    Figure Lengend Snippet: A) Immunofluorescent images of ATDC5 cells on normal gels (upper panel) treated with 30nM GSK101 for 28 hr and severe OA gels with control treatment (lower panel) stained for the nucleus (Hoechst, blue), f-actin (purple), and TRPV4 (white). The left panel shows the composite images of all stains (scale bar =10 μm). The middle panel shows nuclear and f-actin staining, and the right panel shows TRPV4 and nuclear staining. B) TRPV4 fluorescence of ATDC5 cells on normal, OA, and severe OA gels during control (dark blue) and 28 hr of 30 nM GSK101 (light blue) treatments. f-actin was used to draw ROI around the cells to measure TRPV4 fluorescence from max projections of z-stack images. Error Bars are ± SEM, * indicates p-value

    Article Snippet: A subset of cells received 30 nM GSK101 for 28 h. Cells were fixed in 4% paraformaldehyde, blocked for 1 hr in PBS containing 1% BSA/0.2% Fish Gelatin (Fisher Scientific), and probed for membrane TRPV4 using 1:200 rabbit anti-TRPV4 (Alomone Labs, ACC-124) overnight at 4°C.

    Techniques: Staining, Fluorescence

    A) mRNA expression of anabolic factors (Sox-9, aggrecan, col2 and TRPV4) in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. B) mRNA expression of catabolic chondrocyte factors, collagen I and MMP-13 in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. Fold change was normalized to rplp0 expression for each sample and then normalized to the normal control condition. C) Western blotting shows TRPV4 protein levels for ATDC5 cells on normal, OA, and severe OA gels during control treatment. The bar graph represents the normalized TRPV4 protein level to GAPDH for each sample. Error bars are ± SEM, * indicates significance with a p-value

    Journal: bioRxiv

    Article Title: Extracellular Matrix Stiffness Alters TRPV4 Regulation in Chondrocytes

    doi: 10.1101/2021.09.14.460172

    Figure Lengend Snippet: A) mRNA expression of anabolic factors (Sox-9, aggrecan, col2 and TRPV4) in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. B) mRNA expression of catabolic chondrocyte factors, collagen I and MMP-13 in ATDC5 cells on normal, OA and severe OA gels during control (dark blue) or 28 hr of 30 nM GSK101 (light blue) treatment. Fold change was normalized to rplp0 expression for each sample and then normalized to the normal control condition. C) Western blotting shows TRPV4 protein levels for ATDC5 cells on normal, OA, and severe OA gels during control treatment. The bar graph represents the normalized TRPV4 protein level to GAPDH for each sample. Error bars are ± SEM, * indicates significance with a p-value

    Article Snippet: A subset of cells received 30 nM GSK101 for 28 h. Cells were fixed in 4% paraformaldehyde, blocked for 1 hr in PBS containing 1% BSA/0.2% Fish Gelatin (Fisher Scientific), and probed for membrane TRPV4 using 1:200 rabbit anti-TRPV4 (Alomone Labs, ACC-124) overnight at 4°C.

    Techniques: Expressing, Western Blot

    Transfection of large mouse cholangiocytes (MLC) with plasmid for transient receptor potential vanilloid member 4 (TRPV4) small interfering (si)RNA decreases protein and RNA expression. A : representative Western blot demonstrating change in TRPV4 protein levels in cells transfected with nontargeting siRNA (scramble) and cells transfected with TRPV4 siRNA. β-Actin was used as loading control. B : cumulative data demonstrating change in TRPV4 protein ( left ) and mRNA ( right ) levels in control cells, cells transfected with nontargeting siRNA (scramble), and cells transfected with TRPV4 siRNA (* P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Mechanosensor transient receptor potential vanilloid member 4 (TRPV4) regulates mouse cholangiocyte secretion and bile formation

    doi: 10.1152/ajpgi.00176.2019

    Figure Lengend Snippet: Transfection of large mouse cholangiocytes (MLC) with plasmid for transient receptor potential vanilloid member 4 (TRPV4) small interfering (si)RNA decreases protein and RNA expression. A : representative Western blot demonstrating change in TRPV4 protein levels in cells transfected with nontargeting siRNA (scramble) and cells transfected with TRPV4 siRNA. β-Actin was used as loading control. B : cumulative data demonstrating change in TRPV4 protein ( left ) and mRNA ( right ) levels in control cells, cells transfected with nontargeting siRNA (scramble), and cells transfected with TRPV4 siRNA (* P

    Article Snippet: After blocking, immunoblots were incubated overnight with anti-mouse TRPV4 (1:200, Alomone Laboratories) This was followed by incubation with peroxidase-conjugated goat antirabbit antibody (dilution 1:10,000, Jackson Immuno Research Laboratories) and visualized with an ECL+ detection kit (GE Healthcare).

    Techniques: Transfection, Plasmid Preparation, RNA Expression, Western Blot

    Flow-stimulated increases in intracellular calcium concentration ([Ca 2+ ] i ) in large mouse cholangiocytes (MLC). Representative Ca 2+ fluorescence in response to fluid flow. MLC on coverglass were loaded with fura-2 AM and exposed to flow (2 mL/min; shear = 0.64 dyne/cm 2 ); y -axis values represent the ratio of fluorescence at 340 and 380 nm; x -axis represents time (bar = 100 s). Control cells shown ( A ) vs. cells exposed to HC-067047 (HC, 10 µM; B ) and after transfection with transient receptor potential vanilloid member 4 (TRPV4) small interfering (si)RNA ( C ). Cells were exposed to the Ca 2+ ionophore ionomycin, as an internal control. D : cumulative data demonstrating the change in [Ca 2+ ] i in response to fluid flow (shear = 0.64 dyne/cm 2 ) in control cells ( n = 78) and in the presence of HC-067047 (HC, n = 61), or after transfection with TRPV4 siRNA ( n = 49). Values represent change in fluorescence ratio 340/380. Maximal increase vs. basal # P ≤ 0.01; inhibitor HC067047 vs. control ** P ≤ 0.01; * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Mechanosensor transient receptor potential vanilloid member 4 (TRPV4) regulates mouse cholangiocyte secretion and bile formation

    doi: 10.1152/ajpgi.00176.2019

    Figure Lengend Snippet: Flow-stimulated increases in intracellular calcium concentration ([Ca 2+ ] i ) in large mouse cholangiocytes (MLC). Representative Ca 2+ fluorescence in response to fluid flow. MLC on coverglass were loaded with fura-2 AM and exposed to flow (2 mL/min; shear = 0.64 dyne/cm 2 ); y -axis values represent the ratio of fluorescence at 340 and 380 nm; x -axis represents time (bar = 100 s). Control cells shown ( A ) vs. cells exposed to HC-067047 (HC, 10 µM; B ) and after transfection with transient receptor potential vanilloid member 4 (TRPV4) small interfering (si)RNA ( C ). Cells were exposed to the Ca 2+ ionophore ionomycin, as an internal control. D : cumulative data demonstrating the change in [Ca 2+ ] i in response to fluid flow (shear = 0.64 dyne/cm 2 ) in control cells ( n = 78) and in the presence of HC-067047 (HC, n = 61), or after transfection with TRPV4 siRNA ( n = 49). Values represent change in fluorescence ratio 340/380. Maximal increase vs. basal # P ≤ 0.01; inhibitor HC067047 vs. control ** P ≤ 0.01; * P

    Article Snippet: After blocking, immunoblots were incubated overnight with anti-mouse TRPV4 (1:200, Alomone Laboratories) This was followed by incubation with peroxidase-conjugated goat antirabbit antibody (dilution 1:10,000, Jackson Immuno Research Laboratories) and visualized with an ECL+ detection kit (GE Healthcare).

    Techniques: Concentration Assay, Fluorescence, Transfection

    Specific agonists of transient receptor potential vanilloid member 4 (TRPV4) activate currents and increase intracellular calcium concentration ([Ca 2+ ] i ) in large mouse cholangiocytes (MLC). A, top : representative whole cell recording. The specific TRPV4 agonist 4α -phorbol-12,13-didecanoate (4αPDD; 1 µM) activates whole cell currents. Currents measured at +100 mV (○) and at −100 mV (●) are shown. Voltage protocol −100 mV to +100 mV ramp every 5 s. Bottom : cells on coverglass were loaded with fura-2 AM, washed with PBS, and exposed to 4αPDD (1 µM); y -axis values represent the ratio of fluorescence at 340 and at 380 nm. B, top : the specific TRPV4 agonist GSK (100 nM) activates whole cell currents. Currents measured at +100 mv (○) and at −100 mV (●) are shown. Bottom : fura-2-loaded MLC cells were exposed to glycogen synthase kinase (GSK; 100 nM). The y -axis values represent the ratio of fluorescence at 340 and at 380 nm. C, top : cumulative data demonstrating maximal current density (pA/pF) measured at +100 mV in response to 4αPDD or GSK. Currents were significantly inhibited by ruthenium red (RR, 10 µM, n = 9), HC-067047 (HC, 10 µM, n = 16), and after transfection with TRPV4 small interfering (si)RNA ( n = 8). Bottom : cumulative data demonstrating the change in [Ca 2+ ] i in response to 4αPDD or GSK in control cells ( n = 78) and in the presence of RR ( n = 46), HC ( n = 54), or after transfection with TRPV4 siRNA ( n = 27). Values represent relative change in fluorescence ratio 340/380. Maximal increase vs. basal ** P ≤ 0.01, inhibitor vs. control, * P ≤ 0.05, vs. control ** P ≤ 0.01, * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Mechanosensor transient receptor potential vanilloid member 4 (TRPV4) regulates mouse cholangiocyte secretion and bile formation

    doi: 10.1152/ajpgi.00176.2019

    Figure Lengend Snippet: Specific agonists of transient receptor potential vanilloid member 4 (TRPV4) activate currents and increase intracellular calcium concentration ([Ca 2+ ] i ) in large mouse cholangiocytes (MLC). A, top : representative whole cell recording. The specific TRPV4 agonist 4α -phorbol-12,13-didecanoate (4αPDD; 1 µM) activates whole cell currents. Currents measured at +100 mV (○) and at −100 mV (●) are shown. Voltage protocol −100 mV to +100 mV ramp every 5 s. Bottom : cells on coverglass were loaded with fura-2 AM, washed with PBS, and exposed to 4αPDD (1 µM); y -axis values represent the ratio of fluorescence at 340 and at 380 nm. B, top : the specific TRPV4 agonist GSK (100 nM) activates whole cell currents. Currents measured at +100 mv (○) and at −100 mV (●) are shown. Bottom : fura-2-loaded MLC cells were exposed to glycogen synthase kinase (GSK; 100 nM). The y -axis values represent the ratio of fluorescence at 340 and at 380 nm. C, top : cumulative data demonstrating maximal current density (pA/pF) measured at +100 mV in response to 4αPDD or GSK. Currents were significantly inhibited by ruthenium red (RR, 10 µM, n = 9), HC-067047 (HC, 10 µM, n = 16), and after transfection with TRPV4 small interfering (si)RNA ( n = 8). Bottom : cumulative data demonstrating the change in [Ca 2+ ] i in response to 4αPDD or GSK in control cells ( n = 78) and in the presence of RR ( n = 46), HC ( n = 54), or after transfection with TRPV4 siRNA ( n = 27). Values represent relative change in fluorescence ratio 340/380. Maximal increase vs. basal ** P ≤ 0.01, inhibitor vs. control, * P ≤ 0.05, vs. control ** P ≤ 0.01, * P

    Article Snippet: After blocking, immunoblots were incubated overnight with anti-mouse TRPV4 (1:200, Alomone Laboratories) This was followed by incubation with peroxidase-conjugated goat antirabbit antibody (dilution 1:10,000, Jackson Immuno Research Laboratories) and visualized with an ECL+ detection kit (GE Healthcare).

    Techniques: Concentration Assay, Fluorescence, Transfection