Journal: Journal of Inflammation Research

Article Title: Characterization of Early Inflammatory Events Leading to Provoked Vulvodynia Development in Rats

doi: 10.2147/JIR.S367193

Figure Lengend Snippet: TRPV1 and TRPA1 expression in vulvar nerves after multiple rounds of zymosan challenge. ( A ) The expression of TRPV1 channel in vulva nerves, 20 days after the 3rd zymosan/saline administration. Coexpression of TRPV1 channel (green) and neuronal PGP 9.5 (red; White arrows) merged with dapi stain (blue). ( B ) The expression of TRPA1 channel in vulva nerves, 20 days after the 3rd round zymosan\saline administration. Coexpression of TRPA1 channel (red) and neuronal PGP 9.5 (green; White arrows) merged with dapi stain (blue). Scale bar: 50 µm. ( C ) Fluorescence intensity (arbitrary units) of TRPV1, TRPA1, TRPV1/PGP-9.5, and TRPA1/PGP-9.5 in the zymosan and the saline groups 20 days of the 3rd round (n=8 per group). Mean ± SEM. * P <0.05, ** P <0.005, *** P <0.001.

Article Snippet: Afterward, to remove background staining, we used Background Buster (#NB306-50, INNOVEX), followed by three cycles of wash buffer for two minutes each, then incubated for one hour at room temperature in a humidity chamber with one of the first antibodies: anti-rat TRPV1 channel ATTO-488 (1:240, cat.ACC-029-AG, Alomone labs, Jerusalem), anti-rat TRPA1 channel (1:240, cat.ACC-037, Alomone labs, Jerusalem), anti-rat protein gene product 9.5 (PGP 9.5, 1:500, NB300-676, Novus Biological), rabbit anti-Chymase (1:250, NBP2-257441, Novus Biological) and mouse anti-Tryptase (1:250, NBP2-26444, Novus Biological).

Techniques: Expressing, Staining, Fluorescence

Journal: Journal of Inflammation Research

Article Title: Characterization of Early Inflammatory Events Leading to Provoked Vulvodynia Development in Rats

doi: 10.2147/JIR.S367193

Figure Lengend Snippet: TRPV1 and TRPA1 expression in vulvar nerves after 160 days of the 3rd round of zymosan/saline administration. ( A ) The expression of TRPV1 channel in vulva nerves, 20 days after the 3rd zymosan/saline administration. Coexpression of TRPV1 channel (green) and neuronal PGP 9.5 (red; White arrows) merged with dapi stain (blue). ( B ) The expression of TRPA1 channel in vulva nerves, 20 days after the 3rd round zymosan\saline administration. Coexpression of TRPA1 channel (red) and neuronal PGP 9.5 (green; White arrows) merged with dapi stain (blue). Scale bar: 50 µm. ( C ) Fluorescence intensity (arbitrary units) of TRPV1, TRPA1, TRPV1/PGP-9.5, and TRPA1/PGP-9.5 in the zymosan and the saline groups 160 days of the 3rd round (n=8 per group). Mean ± SEM. *** P <0.001.

Article Snippet: Afterward, to remove background staining, we used Background Buster (#NB306-50, INNOVEX), followed by three cycles of wash buffer for two minutes each, then incubated for one hour at room temperature in a humidity chamber with one of the first antibodies: anti-rat TRPV1 channel ATTO-488 (1:240, cat.ACC-029-AG, Alomone labs, Jerusalem), anti-rat TRPA1 channel (1:240, cat.ACC-037, Alomone labs, Jerusalem), anti-rat protein gene product 9.5 (PGP 9.5, 1:500, NB300-676, Novus Biological), rabbit anti-Chymase (1:250, NBP2-257441, Novus Biological) and mouse anti-Tryptase (1:250, NBP2-26444, Novus Biological).

Techniques: Expressing, Staining, Fluorescence

Journal: Journal of Inflammation Research

Article Title: Characterization of Early Inflammatory Events Leading to Provoked Vulvodynia Development in Rats

doi: 10.2147/JIR.S367193

Figure Lengend Snippet: Long-term effects of mast cells stabilizer KF on TRPV1 and TRPA1 expression in vulvar nerves after 81 days of the 3rd round of zymosan/saline administration. ( A ) The expression of TRPV1 channel in vulva nerves, 81 days after the 3rd zymosan/saline administration. Coexpression of TRPV1 channel (green) and neuronal PGP 9.5 (red; white arrows) merged with dapi stain (blue). ( B ) The expression of TRPA1 channel in vulva nerves, 81 days after the 3rd round zymosan\saline administration. Coexpression of TRPA1 channel (red) and neuronal PGP 9.5 (green; white arrows) merged with dapi stain (blue). Scale bar: 50 µm. ( C ) Fluorescence intensity (arbitrary units) of TRPV1, TRPA1, TRPV1/PGP-9.5, and TRPA1/PGP-9.5 in the Saline- ketotifen, the zymosan- ketotifen, and the zymosan-saline group 81 of the 3rd round (n=7 per group). Mean ± SEM. * P <0.05, *** P <0.001.

Article Snippet: Afterward, to remove background staining, we used Background Buster (#NB306-50, INNOVEX), followed by three cycles of wash buffer for two minutes each, then incubated for one hour at room temperature in a humidity chamber with one of the first antibodies: anti-rat TRPV1 channel ATTO-488 (1:240, cat.ACC-029-AG, Alomone labs, Jerusalem), anti-rat TRPA1 channel (1:240, cat.ACC-037, Alomone labs, Jerusalem), anti-rat protein gene product 9.5 (PGP 9.5, 1:500, NB300-676, Novus Biological), rabbit anti-Chymase (1:250, NBP2-257441, Novus Biological) and mouse anti-Tryptase (1:250, NBP2-26444, Novus Biological).

Techniques: Expressing, Staining, Fluorescence

Journal: Medical Oncology (Northwood, London, England)

Article Title: Optimized flow cytometric detection of transient receptor potential vanilloid-1 (TRPV1) in human hematological malignancies

doi: 10.1007/s12032-022-01678-z

Figure Lengend Snippet: TRPV1 gene and protein expression using RNA-sequencing data and Western blotting. A TRPV1 gene expression in the MILE dataset showing an increased TRPV1 expression in patients with different subtypes of ALL compared to normal bone marrow. B Overexpression of TRPV1 gene levels in AML pediatric patients with primary tumor and relapsed disease in the TARGET-AML dataset. All statistical analyses in A and B were performed in comparison to normal healthy subjects reported within each dataset (Dunnett’s multiple comparisons tests). C TRPV1 expression detected using Western blotting in THP-1 and U266B1 cell lines, and PBMCs protein samples of patients with hematological malignancies. THP-1 protein was used as a positive control, and GAPDH was used as an internal control. TRPV1 monomer (~ 95 kDa) was detected in five patients (P#13, P14, P15, P33, P35)

Article Snippet: This was in contrast to the affinity purified anti-TRPV1 antibodies from Alomone Labs and Santa Cruz Biotechnology which were unable to separate the TRPV1 signal from the isotype control, suggesting a lack of specificity for TRPV1.

Techniques: Expressing, RNA Sequencing Assay, Western Blot, Over Expression, Positive Control

Journal: Medical Oncology (Northwood, London, England)

Article Title: Optimized flow cytometric detection of transient receptor potential vanilloid-1 (TRPV1) in human hematological malignancies

doi: 10.1007/s12032-022-01678-z

Figure Lengend Snippet: TRPV1 expression using flow cytometry. A Overview structure of TRPV1 and the 3 antibodies binding sites. Assessment of anti-TRPV1 antibodies: B SC-20813, C ACC-030, and D LS-C150735. Panel D demonstrates clear separation of isotype control from TRPV1 signals in THP-1, U266B1, U937 cells, and healthy human lymphocytes using LS-C150735. E.1 Flow cytometry analysis of TRPV1 expression levels in all patients with hematological malignancies ( n = 49) and healthy controls ( n = 20). Patients were then stratified and TRPV1 expression levels were plotted based on patients’ blood cancer type ( E.2 ) and treatment status ( E.3 ). TRPV1 in MM, B-NHL, and other hematological malignancies group (treated and de novo) were compared to control. Median fluorescence intensity (MFI) value for individual patients expressed using violin plots with the mean and standard deviation is also shown. Overall TRPV1 was significantly higher in all patients with hematological malignancies compared to healthy control ( P = 0.0351, unpaired t test) with B-NHL patients having significantly higher TRPV1 compared to control ( P = 0.0294, Dunnett’s test)

Article Snippet: This was in contrast to the affinity purified anti-TRPV1 antibodies from Alomone Labs and Santa Cruz Biotechnology which were unable to separate the TRPV1 signal from the isotype control, suggesting a lack of specificity for TRPV1.

Techniques: Expressing, Flow Cytometry, Binding Assay, Fluorescence, Standard Deviation

Journal: European Journal of Histochemistry : EJH

Article Title: TRPC1 expression and distribution in rat hearts

doi: 10.4081/ejh.2009.e26

Figure Lengend Snippet: T-PCR based detection of TRPC1 in rat hearts. PCR products were observed in ethidium bromide-stained agarose gel. TRPC1 DNA fragments (467 bp) were amplified from left atrium, right atrium, left ventricle and right ventricle of rats.

Article Snippet: Sections were incubated at 4°C overnight with rabbit anti-rat TRPC1 primary antibodies (1:100 dilution, batch number AN-04, Alomone Labs, Jerusalem, Israel).

Techniques: Staining, Agarose Gel Electrophoresis, Amplification

Journal: European Journal of Histochemistry : EJH

Article Title: TRPC1 expression and distribution in rat hearts

doi: 10.4081/ejh.2009.e26

Figure Lengend Snippet: Immunohistochemical detection of TRPC1 protein in rat hearts. Sections were incubated with primary antibody for TRPC1 (A, B, C, D), without primary antibody (E, F, G, H) or with primary antibody preabsorbed by TRPC1 peptide for negative control (I). Positive signals in brown color can be visualized in the myocytes of the left ventricle (A) and atrium (B), endothelial and smooth muscle layers of coronary arterioles (C), and skeletal muscle cells (D, as positive control). No positive signal could be observed in control experiments without primary antibody. A faint signal was occasionally observed in antigen preabsorption control (I). There are negative cells in the edge of ventricular tissues (J) and also the fibroblasts between ventricular myocytes which showed blue nuclei without positive signals. The right ventricle shows the same distribution of TRPC1 positive signal (K) as the left ventricle. TRPC1 showed intense staining on the cell membranes of ventricular myocytes (A, K, L) and skeletal muscle cells (D). The longitudinal section of left ventricle also shows striated distribution of TRPC1 (L). Scale bar =10 µm, except scale bar = 50 µm in panel J.

Article Snippet: Sections were incubated at 4°C overnight with rabbit anti-rat TRPC1 primary antibodies (1:100 dilution, batch number AN-04, Alomone Labs, Jerusalem, Israel).

Techniques: Immunohistochemical staining, Incubation, Negative Control, Positive Control, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: TRPC1 expression and distribution in rat hearts

doi: 10.4081/ejh.2009.e26

Figure Lengend Snippet: Distribution of TRPC1 in Purkinje cells. These sections were contiguous tissue cross-sections. Endocardial layers were shown. Positive signals, brown in color, could be visualized in Purkinje cells (A, black arrows showed Purkinje cells). The cells beneath the endocardium with loose cytoplasmic structure and without any structure around nuclei were Purkinje cells according to HE staining (B, black arrows showed Purkinje cells). No positive signal could be observed in control experiments (C). Scale bar = 10 µm.

Article Snippet: Sections were incubated at 4°C overnight with rabbit anti-rat TRPC1 primary antibodies (1:100 dilution, batch number AN-04, Alomone Labs, Jerusalem, Israel).

Techniques: Staining

Journal: European Journal of Histochemistry : EJH

Article Title: TRPC1 expression and distribution in rat hearts

doi: 10.4081/ejh.2009.e26

Figure Lengend Snippet: Localization of TRPC1 in rat cardiomyocytes shown by confocal images. Cardiac myocytes were double stained by anti-TRPC1 antibody (A and D) and phalloidin (B and E). Panels C and F show a merged image of panel A/B and D/E respectively, where TRPC1 is colored in red and phalloidin in green. The transverse striation of actin filaments can be seen both in the ventricular myocytes (B) and the atrial myocytes (E). Note that TRPC1 in the ventricular myocyte (A) are parallel to and close to transverse striation of actin filaments, suggesting that they are located at T-tubules while TRPC1 in the atrial myocytes (D) do not show the striation-like distribution. Scale bar =25 µm.

Article Snippet: Sections were incubated at 4°C overnight with rabbit anti-rat TRPC1 primary antibodies (1:100 dilution, batch number AN-04, Alomone Labs, Jerusalem, Israel).

Techniques: Staining