Journal: BIOCELL

Article Title: Nicotinic acid induces apoptosis of glioma cells via the calcium-dependent endoplasmic reticulum stress pathway

doi: 10.32604/biocell.2022.017383

Figure Lengend Snippet: FIGURE 6. TRPV1 mediates NA function in U251 cells. (A–D) Evaluation of TRPV1 knockdown efficiency. U251 cells were transfected with two siRNAs targeting TRPV1, and RT-PCR (A) and immunofluorescence (B–D, pictures were taken with a magnification of 40×) were carried out to detect TRPV1 mRNA and protein levels, respectively. NC, negative control without reverse transcriptase. (E) Time-lapse curve of intracellular [Ca2+] after the transfection of TRPV1 siRNA2 and treatment with 14 mM NA. (F–I) Apoptosis of U251 cells after transfection with TRPV1 siRNA2 and treatment with 14 mM NA. Cells were harvested, stained with annexin-V and PI, and analyzed by flow cytometry. The percentage of U251 cells that underwent apoptosis with NA treatment from three independent experiments is summarized in J (****P < 0.0001).

Article Snippet: The primary antibodies used were cleaved caspase-3 (#9664, 1:200; Cell Signaling Technology) and TRPV1 (AF3066-SP, 1:200; R&D Systems).

Techniques: Knockdown, Transfection, Reverse Transcription Polymerase Chain Reaction, Negative Control, Reverse Transcription, Staining, Cytometry

Journal: BIOCELL

Article Title: Nicotinic acid induces apoptosis of glioma cells via the calcium-dependent endoplasmic reticulum stress pathway

doi: 10.32604/biocell.2022.017383

Figure Lengend Snippet: FIGURE 7. NA induces ER-mediated apoptosis by activating calpains. (A) NA activates calpains in U251 cells. After the indicated treatment, cellular calpain activities were assessed as described in the Materials and Methods section. (B–E) Apoptosis of U251 cells was evaluated by flow cytometry after the treatment with 14 mM NA and indicated inhibitors. The percentage of U251 cells that underwent apoptosis with NA treatment from three independent experiments is summarized in F (**P < 0.01 and ***P < 0.001). (G and H) Our models of NA function in inducing ER stress and apoptosis via TRPV1 and calpains. See the full text for a detailed explanation.

Article Snippet: The primary antibodies used were cleaved caspase-3 (#9664, 1:200; Cell Signaling Technology) and TRPV1 (AF3066-SP, 1:200; R&D Systems).

Techniques: Cytometry

Journal: Journal of nutritional science and vitaminology

Article Title: Effect of Oleuropein on Anti-Obesity and Uncoupling Protein 1 Level in Brown Adipose Tissue in Mild Treadmill Walking Rats with Diet-Induced Obesity.

doi: 10.3177/jnsv.70.193

Figure Lengend Snippet: Fig. 4. Protein levels of transient receptor potential ankyrin subtype 1 (TRPA1) (A), transient receptor potential vanilloid subtype 1 (TRPV1) (B), and brain-derived neurotrophic factor (BDNF) (C) and the results of Western blot (D) in the brain, plasma BDNF level (E), and BDNF (F) and peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (PGC1- ) protein level in the gastrocnemius muscle (G) of rats fed with a high-fat diet (HF) or HF supplemented with 0.08% oleuro- pein (HFO) combined with mild treadmill walking (MTW) for 28 d. Values are expressed as meansSE, n 6 or 7. Means labeled with different letters indicate a signifi cant difference (p0.05). The effects of oleuropein supplementation (O) and MTW (W) and the interaction between O and W (OW) were both signifi cant (p0.05).

Article Snippet: The primary antibodies were as follows: BDNF (15 kDa, ab108319, 1 : 1,000; Abcam), TRPV1 (100 kDa, NBP1-97417, 1 : 1,000; Novus Bio), TRPA1 (100 kDa, NB11040763, 1 : 1,000; Novus Bio), peroxisome proliferatoractivated receptor-gamma coactivator 1-alpha (PGC1; 100 kDa, ab106814, 1 : 1,000; Abcam), -actin (42 kDa, ab185058, 1 : 10,000; Abcam), and -actin (42 kDa, ab20272, 1 : 5,000; Abcam).

Techniques: Derivative Assay, Western Blot, Clinical Proteomics, Labeling

Journal: Molecular Medicine Reports

Article Title: Immunolocalization of cannabinoid receptor type 1 and CB2 cannabinoid receptors, and transient receptor potential vanilloid channels in pterygium

doi: 10.3892/mmr.2017.7246

Figure Lengend Snippet: TRPV immunolocalization in pterygium and conjunctival tissue samples. Cytoplasmic TRPV1 expression in (A) primary pterygium and (B) conjunctiva samples. Endothelial cells are TRPV1 immunopositive. (B, insert) Immunostaining is absent in negative controls sections. (C) Photomicrographs from the same recurrent pterygium sample demonstrating immunostaining heterogeneity for TRPV3 expression in (top) epithelial cells and (bottom) epithelial cells with strong TRPV3 cytoplasmic immunoexpression with wide distribution. Endothelial cells exhibit TRPV3 immonopositivity. Immunostaining was weak for CB1 (LI=5) and absent for CB2 (LI=0) cannabinoid receptors. Photomicrographs from the same primary pterygium sample displaying (D) weak TRPV2 (LI=8) and (E) predominantly nuclear TRPV4 (LI=80) immunoreactivity in epithelial cells. Note that TRPV4 expression is visible in all layers of the pterygium sample, including a few goblet cells. Vessels demonstrate TRPV2 and TRPV4 immunoreactivity. In addition, immunostaining for CB1 (LI=60) and CB2 (LI=50) cannabinoid receptors was detected. (F) TRPV4 immunostaining is localized in the cytoplasm and nucleus of basal and suprabasal epithelial cells, as well as in certain lamina propria cells in this conjunctival sample. Counterstain, hematoxylin; original magnification, ×400; scale bar, 50 µm. TRPV, transient receptor potential cation channel subfamily V member; CB1, cannabinoid receptor type 1; LI, labeling index; CB2, cannabinoid receptor type 2.

Article Snippet: TRPV channel expression was detected using polyclonal rabbit anti-TRPV1 antibody (cat no. NBP1-71774; dilution, 1:200) manufactured by Novus Biologicals, Ltd. (Cambridge, UK), polyclonal rabbit anti-TRPV2 (cat no. TA317464; dilution 1:200) and monoclonal mouse anti-TRPV3 antibody (cat no. AM20072PU-N; dilution 1:300) produced by Acris Antibodies GmbH (Herford, Germany) and rabbit polyclonal anti-TRPV4 antibody from Abcam (cat no. ab39260; dilution 1:200 Cambridge, UK).

Techniques: Expressing, Immunostaining, Labeling

Journal: Molecular Medicine Reports

Article Title: Immunolocalization of cannabinoid receptor type 1 and CB2 cannabinoid receptors, and transient receptor potential vanilloid channels in pterygium

doi: 10.3892/mmr.2017.7246

Figure Lengend Snippet: Immunohistochemical expression of TRPV channels in human conjunctiva and pterygium epithelium.

Article Snippet: TRPV channel expression was detected using polyclonal rabbit anti-TRPV1 antibody (cat no. NBP1-71774; dilution, 1:200) manufactured by Novus Biologicals, Ltd. (Cambridge, UK), polyclonal rabbit anti-TRPV2 (cat no. TA317464; dilution 1:200) and monoclonal mouse anti-TRPV3 antibody (cat no. AM20072PU-N; dilution 1:300) produced by Acris Antibodies GmbH (Herford, Germany) and rabbit polyclonal anti-TRPV4 antibody from Abcam (cat no. ab39260; dilution 1:200 Cambridge, UK).

Techniques: Immunohistochemical staining, Expressing, Standard Deviation

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: List of antibodies.

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Purification, Muscles, Clinical Proteomics

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Characterization of the TRPV1 antibody in the monkey retina. Western blot analysis of total protein samples from a vervet monkey retina, showing detection of the expected protein band at 100 kDa ( A ). The band was not detected when the antibody was pre-incubated with its corresponding TRPV1 blocking peptide (BP) at the ratio 1:10 ( A ). All lanes contained 10 μg of total protein. Immunohistochemistry on vervet retinal tissue with the antibody raised against TRPV1 revealed a unique staining profile ( B ). When the TRPV1 antibody was pre-incubated with its corresponding BP, it revealed an absence of staining in the vervet retinas ( C ). Quantitative analysis of the TRPV1 fluorescent signal through the six different retinal layers reveals a relatively higher signal intensity in the OPL and the GCL layers compared to the photoreceptor (PRL) and the outer nuclear (ONL) layers (P < 0.05) ( D ). TRPV1 transient receptor potential vanilloid type 1, BP blocking peptide, LC loading control, PRL photoreceptor layer, ONL outer nuclear layer, OPL outer plexiform layer, INL inner nuclear layer, IPL inner plexiform layer, GCL ganglion cell layer. Scale bar = 75 μm. Mean optical density ratios ± standard error of the mean (SEM) are depicted.

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Western Blot, Incubation, Blocking Assay, Immunohistochemistry, Staining, Control

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Labeling pattern of TRPV1-IR throughout the vervet retina. Confocal micrographs taken from the center ( A ), middle ( B ), and peripheral retina ( C ). Quantitative analysis of the TRPV1 fluorescent signal through the three distinct retinal eccentricities shows relatively higher central signal intensity in the IPL and the RGC layers, and greater peripherical labeling in the OPL and the ONL layers (P < 0.05) ( D ). PRL photoreceptor layer, ONL outer nuclear layer, OPL outer plexiform layer, INL inner nuclear layer, IPL inner plexiform layer, GCL ganglion cell layer, GCFL ganglion cell fiber layer. Scale bar = 75 µm. Mean optical density ratios ± standard error of the mean (SEM) are depicted.

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Labeling

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Double-label immunofluorescence illustrating immunolabeling of TRPV1 in ganglion cells, horizontal cells, and amacrine cells. ( A-C ) The antibody against Brn3a labels the nucleus of ganglion cells in the monkey retina, and these cells were strongly TRPV1 immunoreactive. Arrows indicate Brn3a-positive ganglion cells that are TRPV1 immunoreactive. Each protein expression is presented alone in grayscale: Brn3a ( A ) and TRPV1 ( B ); then, the two are presented merged ( C ), Brn3a in green and TRPV1 in red). ( D-I ) TRPV1 is expressed in horizontal cell synaptic terminals (arrows in OPL) and cell bodies (arrowheads in the INL). Each expression is presented alone in grayscale: PV ( D,G ), TRPV1 ( E,H ); then, the two are presented merged ( F,J ). ( J – L ) Arrows show the expression of TRPV1 in the amacrine cell bodies (INL). Each expression is presented alone in grayscale: syntaxin-positive amacrine cells ( J ) and TRPV1 ( K ); then, the two are presented merged ( L ). INL inner nuclear layer, IPL inner plexiform layer, OPL outer plexiform layer, GCL ganglion cell layer. Scale = 75 μm for ( A – C ) and ( J – L ), 30 μm for ( D – F ) and 10 μm for ( G – I ).

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Immunofluorescence, Immunolabeling, Expressing

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Double-label immunofluorescence showing no signal for TRPV1 in rod bipolar cells and Müller cells. TRPV1 is neither expressed in the cell bodies (small arrows in the upper INL) of bipolar cells nor their synaptic terminals at the edge of the OPL (arrowheads). Each protein expression is presented alone in grayscale: PKC ( A ) and TRPV1 ( B ); then, the two are presented merged ( C ), PKC in green and TRPV1 in red. Long arrows show specifically that the Müller cell bodies do not express TRPV1. Each protein expression is presented alone in grayscale: glutamine synthetase ( D ) and TRPV1 ( E ); then, the two are presented merged ( F ), glutamine synthetase in green and TRPV1 in red. Scale = 75 μm for ( A – F ).

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Immunofluorescence, Expressing

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Mapping of the receptors CB1R, CB2R, GPR55, and TRPV1 in the monkey retina. These receptors are differently expressed in the retina of monkeys. These results are compiled from several published articles , , . CB1R is represented in green, CB2R in magenta, GPR55 in blue, and TRPV1 in red. OS, photoreceptors outer segments; IS, photoreceptors inner segments; ONL, outer nuclear layer; ONL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques:

Journal: Scientific Reports

Article Title: Transient receptor potential vanilloid type 1 is expressed in the horizontal pathway of the vervet monkey retina

doi: 10.1038/s41598-020-68937-9

Figure Lengend Snippet: Schematic illustration of the hypothetical mechanism of action of TRPV1 in horizontal cells of the monkey retina following stimulation of a cone. The different steps are highlighted by numbers. (1) When a cone is stimulated (C1), endocannabinoids (eCBs), particularly AEA and 2-AG, are produced in the synaptic cleft. (2) TRPV1 receptors are activated by eCBs in horizontal cells. (3) The calcium influx through the membrane causes depolarization of the horizontal cell. (4) GABA release in the synaptic cleft. (5) GABA binds to GABA receptors on the presynaptic neighboring cone pedicle (5). This mechanism of lateral inhibition could also potentially modulate the information transmitted to the bipolar cells and retinal ganglion cells. S spot stimulus, C1 stimulated cone 1, C2 adjacent cone 2, H horizontal cell, eCB endocannabinoid, GABAr Gamma-amino-butyric acid receptors, TRPV1 Transient receptor potential vanilloid type 1.

Article Snippet: For the control condition, the same protocol was ran simultaneously as described, excepting that the anti-TRPV1 primary antibody was preincubated with its blocking peptide (BP; #NB100-1617PEP, Novus Biologicals, Littleton, CO, USA) in a 1:10 dilution for 1 h.

Techniques: Produced, Membrane, Inhibition

Journal: BPB Reports

Article Title: Involvement of TRPV1 and TRPV4 Channels in Enhancement of Metastatic Ability Induced by γ-Irradiation in Human Lung Cancer A549 Cells

doi: 10.1248/bpbreports.3.1_50

Figure Lengend Snippet: Fig. 1. Effect of TRPV1 and TRPV4 Inhibitors on γ-Irradiation-Induced Migration of A549 Cells

Article Snippet: The primary antibodies used were anti-β-actin monoclonal antibody (FUJIFILM Wako Pure Chemical, Osaka, Japan), TRPV1 antibody (Novus), and TRPV4 antibody (Sigma Aldrich, USA).

Techniques: Irradiation, Migration

Journal: BPB Reports

Article Title: Involvement of TRPV1 and TRPV4 Channels in Enhancement of Metastatic Ability Induced by γ-Irradiation in Human Lung Cancer A549 Cells

doi: 10.1248/bpbreports.3.1_50

Figure Lengend Snippet: Fig. 2. Effect of TRPV1 and TRPV4 Inhibitors on γ-Irradiation-Induced Actin Remodeling in A549 Cells

Article Snippet: The primary antibodies used were anti-β-actin monoclonal antibody (FUJIFILM Wako Pure Chemical, Osaka, Japan), TRPV1 antibody (Novus), and TRPV4 antibody (Sigma Aldrich, USA).

Techniques: Irradiation

Journal: BPB Reports

Article Title: Involvement of TRPV1 and TRPV4 Channels in Enhancement of Metastatic Ability Induced by γ-Irradiation in Human Lung Cancer A549 Cells

doi: 10.1248/bpbreports.3.1_50

Figure Lengend Snippet: Fig. 4. Effect of TRPV1 and TRPV4 Channel Inhibitors on γ-Irradiation- Induced B16 Melanoma Cell Metastasis to the Lungs

Article Snippet: The primary antibodies used were anti-β-actin monoclonal antibody (FUJIFILM Wako Pure Chemical, Osaka, Japan), TRPV1 antibody (Novus), and TRPV4 antibody (Sigma Aldrich, USA).

Techniques: Irradiation