Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Detection of TRPV4 expression in gastric adenocarcinoma tissues and adjacent noncancerous tissues by IHC staining of tissue microarrays. ( A ) Representative images of TRPV4 expression in gastric adenocarcinoma tissues and adjacent noncancerous tissues. (a, b) Weakly focal positive TRPV4 expression in adjacent noncancerous gastric epithelia. (c, d) Strongly positive TRPV4 expression in a case of gastric adenocarcinoma. ( B ) Scatterplots of IHC scores of TRPV4 expression in gastric adenocarcinoma tissues and adjacent noncancerous tissues. ( C ) Percents of tissue samples with high TRPV4 expression and low TRPV4 expression based on the cut-off value of IHC scores in gastric adenocarcinoma tissues and adjacent noncancerous tissues (***P<0.001). Magnification: (a,c) ×50 and (b,d) ×400. Abbreviations: IHC, immunohistochemical; TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Association of TRPV4 Expression with Clinicopathological Characteristics in Patients with Gastric Cancer

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Representation images of TRPV4, E-cadherin and vimentin expression in gastric cancer tissues by immunohistochemical staining. Magnification, ×400. ( A – C ) High TRPV4 expression in a case of poor differentiated gastric adenocarcinoma with loss expression of E-cadherin and positive vimentin expression. ( D – F ) Low TRPV4 expression in a case of signet ring cell carcinoma with high E-cadherin expression and negative Vimentin expression. Abbreviation: TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Correlation Between Expression of TRPV4, E-Cadherin and Vimentin in Gastric Cancer

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Survival curves using the Kaplan–Meier method by Log rank test. ( A and B ) The patients with high TRPV4 expression had shorter overall survival and disease-free survival than those with low TRPV4 expression. ( C and D ) The patients with low E-cadherin expression had shorter overall survival and disease-free survival than those with high E-cadherin expression. ( E and F ) The patients with high vimentin expression had shorter overall survival and disease-free survival than those with low vimentin expression. ( G ) Overall survival analysis of TRPV4 gene in gastric cancer obtained from Kaplan–Meier Plotter online ( http://kmplot.com/analysis/ ). *P<0.05, **P<0.01, ***P<0.001. Abbreviations: Cum, cumulative; TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Univariate and Multivariate Cox Regression Analyses of Overall Survival in Patients with Gastric Cancer

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Univariate and Multivariate Cox Regression Analyses of Disease-Free Survival in Patients with Gastric Cancer

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: The protein expression of TRPV4 in gastric cell lines and down-regulation of TPRV4 inhibited proliferation of gastric cancer cells. ( A ) The protein expression levels of TPRV4 were detected in gastric cancer cell lines (HGC-27 and MGC-803) and normal gastric mucosa cell GES-1 by Western blot. **P<0.01, ***P<0.001. ( B ) Western blot analyses of the expression levels of TRPV4 protein in gastric cancer cell lines following transfection with shTRPV4 or shCtrl. ***P<0.001. ( C ) Proliferation capacities of gastric cancer cells lines were assayed at 24 h, 48 h and 72 h by CCK-8 following transfection with shTRPV4 or shCtrl. ***P<0.001. ( D ) Colony formation assays were performed in gastric cancer cell lines following transfection with shTRPV4 or shCtrl. *P<0.05. Abbreviations: CCK-8, Cell Counting Kit-8; TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing, Western Blot, Transfection, CCK-8 Assay, Cell Counting

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Transwell assays demonstrated that TRPV4 down-regulation greatly inhibited invasion abilities of gastric cancer cells. ( A and B ) Representative images of invasive cells following transfection with shTRPV4 or shCtrl. Magnification, ×200. ( C and D ) The column diagram of the numbers of invasive cells following transfection with shTRPV4 or shCtrl. **P<0.01, ***P<0.001. Abbreviation: TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Transfection

Journal: OncoTargets and therapy

Article Title: TRPV4 Overexpression Promotes Metastasis Through Epithelial–Mesenchymal Transition in Gastric Cancer and Correlates with Poor Prognosis

doi: 10.2147/OTT.S256918

Figure Lengend Snippet: Down-regulation of TPRV4 regulated the expression levels of EMT-related proteins in gastric cancer cell lines ( A ) The expression levels of EMT-related proteins (E-cadherin, vimentin, N-Cadherin, Snail, Slug and Twist) were detected by Western blot assays following transfection with shTRPV4 or shCtrl. ( B ) The column diagram of the expression levels of EMT-related proteins in HGC-27 following transfection with shTRPV4 or shCtrl. ( C ) The column diagram of the expression levels of EMT-related proteins in MGC-803 following transfection with shTRPV4 or shCtrl. *P<0.05, ***P<0.001. Abbreviations: ns, non-significant; EMT, epithelial–mesenchymal transition; TRPV4, transient receptor potential vanilloid 4.

Article Snippet: After blocking with 20% bovine serum albumin (BSA), these pretreated slides were incubated at 4°C overnight with antibodies against TRPV4 (1:100 dilution, #ACC-034-AO, Alomone labs), E-cadherin (1:50 dilution, clone NCH-38, Dako), and vimentin (1:100 dilution, clone V9, Dako), followed by incubation with the HRP-conjugated secondary antibody (Dako).

Techniques: Expressing, Western Blot, Transfection

Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: Primer sequences for reverse transcription-quantitative PCR.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: A HSD in vivo impaired barrier function of bladder. (A) H&E staining and histological score of bladder tissues (scale bar, 1 mm). Data were presented as the median with interquartile range. (B) Bladder weight/body weight ratio. (C) Thickness of lamina propria and mucosal layer of the bladder. Relative mRNA expression levels of (D) inflammatory response markers, IL-1β and TNF-α, (E) tight junction proteins, TJP-1 and Claudin-1 and (F) TRPV4. (G) Representative images of histological staining and quantification of protein expression of TRPV4, TJP-1 and CLAUDIN-1 in bladder tissues sections from CON and HSD mice. Scale bar, 250 µm (n=3). (H) Correlation analysis between the mRNA expression levels of tight junction proteins and inflammation factors in the bladder and urination characteristics in CON and HSD mice. Data are presented as mean ± SD (n=6). P-values were calculated using a two-tailed unpaired Student's t test; *** P<0.001, ** P<0.01 and * P<0.05. HSD, high salt diet; CON, control; TJP-1, tight junction protein 1; TRPV4, transient receptor potential vanilloid 4; ns, non-significant; A.U., arbitrary units.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: In Vivo, Staining, Expressing, Two Tailed Test, Control

Journal: Experimental and Therapeutic Medicine

Article Title: A high salt diet impairs the bladder epithelial barrier and activates the NLRP3 and NF‑κB signaling pathways to induce an overactive bladder in vivo

doi: 10.3892/etm.2024.12651

Figure Lengend Snippet: A HSD increased uroepithelial oxidative stress in SV-HUC-1 cells. Relative mRNA expression levels of (A) TJP-1 and CLAUDIN-1, (B) TRPV4 and (C) IL-1β and TNF-α in HSD-treated and CON cells. (D) Relative MPO expression levels in CON and HSD groups. (E) Representative images and quantification of intracellular ROS levels (scale bar, 200 µm). (F) Relative mRNA expression levels of NCF-1 and CYBA. (G) Relative MDA expression levels in CON and HSD groups. Data are presented as mean ± SD (n=6). P-values were calculated using a two-tailed unpaired Student's t-test; *** P<0.001 and ** P<0.01. HSD, high salt diet; CON, control; SV-HUC-1, SV40 virus transformed human uroepithelium cells; TJP-1, tight junction protein 1; TRPV4, transient receptor potential vanilloid 4; MPO, myeloperoxidase; ROS, reactive oxygen species; NCF-1, neutrophil cytosolic factor 1; CYBA, cytochrome B-245 alpha chain; MDA, malondialdehyde.

Article Snippet: The samples were then incubated at 4˚C overnight with the following primary antibodies: Rabbit anti-TJP-1 antibody (1:250; cat. no. ab276131; Abcam), rabbit anti-CLAUDIN-1 antibody (1:200; cat. no. YT0942; ImmunoWay Biotechnology Company), rabbit anti-TRPV4 antibody (1:200; cat. no. YT5833; ImmunoWay Biotechnology Company), rabbit anti-CASP1 antibody (1:200; cat. no. YP0749; ImmunoWay Biotechnology Company) and rabbit anti-NF-κB antibody (1:200; cat. no. YM8001; ImmunoWay Biotechnology Company).

Techniques: Expressing, Two Tailed Test, Control, Virus, Transformation Assay

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Quantification of TRPV4 expression. (a) Sites where tissue samples were taken and the quantification method for stained areas. Tissue was collected from 57 patients who underwent gastric resection. The following areas were sectioned in all layers: 1. greater curvature of the fornix; 2. greater curvature of the gastric body; 3. greater curvature of the antrum; 4. anterior wall of the gastric body; 5, posterior wall of the gastric body. (b) TRPV4 immunostaining in gastric tissue. (c) The images were captured using a microscope with BZ-X710 (Keyence, Osaka, Japan), and staining was quantified with hybrid cell count BZ-H4C.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Expressing, Staining, Immunostaining, Microscopy, Cell Counting

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Comparison of stained areas between nonobese and obese cases. (a) Renal tissues were used as a positive control for TRPV4-immunostaining. (b) For the negative control, tissues were subjected to immunostaining without the primary antibody. (c) Immunostaining of the nonobese and obese stomach. (d) Comparison of stained areas in tissue samples collected from patients with and without obesity. Lines within the boxes represent median values; upper and lower lines of the boxes represent 25th and 75th percentiles, respectively; upper and lower bars outside the boxes represent the maximum and minimum, respectively ( ∗∗ p < 0.01).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Comparison, Staining, Positive Control, Immunostaining, Negative Control

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: TRPV4 immunostaining in the stomach of patients with and without obesity.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Immunostaining

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Fluorescence immunostaining of TRPV4 in MGN3-1 cells. TRPV4 protein expression was observed in MGN3-1 cells. Magnification: ×600. TRPV4-mediated change in [Ca 2+ ] i in MGN3-1 cells.

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Fluorescence, Immunostaining, Expressing

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: TRPV4-mediated changes in cytosolic Ca 2+ in MGN3-1 cells. The effects of GSK1016790 A and HC 067047 on [Ca 2+ ] i in MGN3-1 cells. Compared to the control level at 180 s, [Ca2+] i increased in MGN3-1 cells treated with GSK1016790 A ( n = 20, p < 0.01) but decreased in cells treated with GSK1016790 A and HC067047 ( n = 20, p < 0.01) (GSK1016790 A, TRPV4 agonist; HC067047, TRPV4 antagonist).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: Control

Journal: International Journal of Endocrinology

Article Title: Potential Role of TRPV4 in Stretch-Induced Ghrelin Secretion and Obesity

doi: 10.1155/2022/7241275

Figure Lengend Snippet: Regulation of ghrelin secretion by the TRPV4 agonist and antagonist in MGN3-1 cells. (a) The amount of secreted acyl ghrelin after four-hour treatment of MGN3-1 cells with GSK1016790 A (3 μ M) or HC067047 (20 μ M) + GSK1016790 A (3 μ M). (b) The amount of secreted des-acyl ghrelin after four-hour treatment of MGN3-1 cells with GSK1016790 A (3 μ M) or HC067047 (20 μ M) + GSK1016790 A (3 μ M) ( ∗ p < 0.05; ∗∗ p < 0.01; n = 6; error bars: standard error of the mean).

Article Snippet: The cells were then probed with a rabbit polyclonal anti-TRPV4 antibody (OST00265 G, Osenses Pty Ltd, Keswick, Australia) overnight at 4°C.

Techniques: