trpv6 antibody  (Alomone Labs)


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    Alomone Labs trpv6 antibody
    Localisation of <t>TRPV6</t> protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).
    Trpv6 Antibody, 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/trpv6 antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trpv6 antibody - by Bioz Stars, 2022-01
    93/100 stars

    Images

    1) Product Images from "TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy"

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    Journal: Reproductive Biology and Endocrinology : RB & E

    doi: 10.1186/1477-7827-10-66

    Localisation of TRPV6 protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).
    Figure Legend Snippet: Localisation of TRPV6 protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).

    Techniques Used: Labeling

    Uterine and placentomal TRPV6 and CaBP-9k mRNA expression. TRPV6 and CaBP-9k mRNA expression in the intercaruncular wall (Figure 1A, C) and in the placenta (n ≥ 3 animals/month) (Figure 1B, D) from the 2 nd to 9 th months of pregnancy, determined by real-time PCR. Bars with ** differ significantly (p
    Figure Legend Snippet: Uterine and placentomal TRPV6 and CaBP-9k mRNA expression. TRPV6 and CaBP-9k mRNA expression in the intercaruncular wall (Figure 1A, C) and in the placenta (n ≥ 3 animals/month) (Figure 1B, D) from the 2 nd to 9 th months of pregnancy, determined by real-time PCR. Bars with ** differ significantly (p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Western blot analyses of placental TRPV6 and CaBP-9k postpartum in cows with (RFM) and without retained fetal membranes (DFM). A) Representative Western blot of TRPV6 in the placenta. Bands of approximately 75 kDa were detected in all animals. B) Relative expression of TRPV6 was determined by measuring the optical density relative to GAPDH using ImageJ software. Significant changes in expression levels were not detected (p > 0.05). C) Representative Western blot of CaBP-9k. D) The relative expression levels to GAPDH were determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.
    Figure Legend Snippet: Western blot analyses of placental TRPV6 and CaBP-9k postpartum in cows with (RFM) and without retained fetal membranes (DFM). A) Representative Western blot of TRPV6 in the placenta. Bands of approximately 75 kDa were detected in all animals. B) Relative expression of TRPV6 was determined by measuring the optical density relative to GAPDH using ImageJ software. Significant changes in expression levels were not detected (p > 0.05). C) Representative Western blot of CaBP-9k. D) The relative expression levels to GAPDH were determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Techniques Used: Western Blot, Expressing, Software, One-tailed Test

    Immunolabelling of CaBP-9k in placentomes and adherent fetal membranes. Localisation of CaBP-9k during pregnancy with strong signals in the inner layer (IL) of the place tome and in the fetal membranes (FM) compared to the outer labyrinth layer (OL) (A, C). Strong staining in the binucleate trophoblast cells (BNC) and in the maternal epithelium (ME) inside the place tome (B, C). Figure 2 F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.
    Figure Legend Snippet: Immunolabelling of CaBP-9k in placentomes and adherent fetal membranes. Localisation of CaBP-9k during pregnancy with strong signals in the inner layer (IL) of the place tome and in the fetal membranes (FM) compared to the outer labyrinth layer (OL) (A, C). Strong staining in the binucleate trophoblast cells (BNC) and in the maternal epithelium (ME) inside the place tome (B, C). Figure 2 F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Techniques Used: Staining, Negative Control

    Immunolabelling of TRPV6 in the placentomes and adherent fetal membranes. Localisation of TRPV6 during pregnancy in the placenta and adherent membranes with strong staining in the inner layer (IL) of the place tome (A, D) and the adherent fetal membranes (FM) (A, C). Considerably weaker staining in the middle (ML) and outer labyrinth (OL) layers (A, B). Scale bars indicate 200 μm.
    Figure Legend Snippet: Immunolabelling of TRPV6 in the placentomes and adherent fetal membranes. Localisation of TRPV6 during pregnancy in the placenta and adherent membranes with strong staining in the inner layer (IL) of the place tome (A, D) and the adherent fetal membranes (FM) (A, C). Considerably weaker staining in the middle (ML) and outer labyrinth (OL) layers (A, B). Scale bars indicate 200 μm.

    Techniques Used: Staining

    Immunolabelling of TRPV6 in the uterine endometrium. Localisation of uterine TRPV6 during pregnancy with strong labelling in the glandular (GE) and luminal epithelia (LE). Figure 2F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.
    Figure Legend Snippet: Immunolabelling of TRPV6 in the uterine endometrium. Localisation of uterine TRPV6 during pregnancy with strong labelling in the glandular (GE) and luminal epithelia (LE). Figure 2F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Techniques Used: Negative Control

    Placental TRPV6 and CaBP-9k mRNA expression post partum in cows with and without retained fetal membranes. Detection of TRPV6 (A) and Calbindin-D9k (CaBP-9k) (B) mRNA levels in the placenta of postpartum cows that retained the fetal membranes for more than 12 hours (RFM; n = 5) and cows that discharged the fetal membranes (DFM; n = 6), measured by real-time PCR (Taqman). No significant changes were observed (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.
    Figure Legend Snippet: Placental TRPV6 and CaBP-9k mRNA expression post partum in cows with and without retained fetal membranes. Detection of TRPV6 (A) and Calbindin-D9k (CaBP-9k) (B) mRNA levels in the placenta of postpartum cows that retained the fetal membranes for more than 12 hours (RFM; n = 5) and cows that discharged the fetal membranes (DFM; n = 6), measured by real-time PCR (Taqman). No significant changes were observed (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, One-tailed Test

    Western blot analyses of endometrial and placentomal TRPV6 protein during pregnancy. A) Representative Western blot of TRPV6 protein expression in the uterine wall (n = 3 animals/month). Bands of approximately 75 kDa were detected. An additional band of approximately 90 kDa was seen in the 2 nd and 3 rd months. After incubation with N-glycosidase F (PNGase F), only the smaller band of 75 kDa was seen, demonstrating that the other band is the glycosylated variant of TRPV6. As a negative control, samples were preincubated with a control antigen. B) Relative expression of TRPV6 to GAPDH was determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). C) Representative Western blot analyses of TRPV6 in the placenta from 2 nd to 9 th months of pregnancy (n = 3 animals/month). D) The expression of TRPV6 relative to GAPDH was determined by measuring the optical density using ImageJ software. No significant changes (p > 0.05) were detected. Data were analysed using a parametric one-way analysis of variance (ANOVA), followed by Dunnett’s comparison test, where the second month was taken as the control group, and are presented as means ± SD.
    Figure Legend Snippet: Western blot analyses of endometrial and placentomal TRPV6 protein during pregnancy. A) Representative Western blot of TRPV6 protein expression in the uterine wall (n = 3 animals/month). Bands of approximately 75 kDa were detected. An additional band of approximately 90 kDa was seen in the 2 nd and 3 rd months. After incubation with N-glycosidase F (PNGase F), only the smaller band of 75 kDa was seen, demonstrating that the other band is the glycosylated variant of TRPV6. As a negative control, samples were preincubated with a control antigen. B) Relative expression of TRPV6 to GAPDH was determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). C) Representative Western blot analyses of TRPV6 in the placenta from 2 nd to 9 th months of pregnancy (n = 3 animals/month). D) The expression of TRPV6 relative to GAPDH was determined by measuring the optical density using ImageJ software. No significant changes (p > 0.05) were detected. Data were analysed using a parametric one-way analysis of variance (ANOVA), followed by Dunnett’s comparison test, where the second month was taken as the control group, and are presented as means ± SD.

    Techniques Used: Western Blot, Expressing, Incubation, Variant Assay, Negative Control, Software

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    Alomone Labs rabbit anti trpv5
    Channel characteristics of wild-type and mutant <t>TRPV5.</t> ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p
    Rabbit Anti Trpv5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti trpv5/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti trpv5 - by Bioz Stars, 2022-01
    92/100 stars
      Buy from Supplier

    93
    Alomone Labs trpv6 antibody
    Localisation of <t>TRPV6</t> protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).
    Trpv6 Antibody, 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/trpv6 antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trpv6 antibody - by Bioz Stars, 2022-01
    93/100 stars
      Buy from Supplier

    Image Search Results


    Channel characteristics of wild-type and mutant TRPV5. ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Channel characteristics of wild-type and mutant TRPV5. ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels [62] . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Mutagenesis, Injection, Mass Spectrometry, Inhibition, Transfection, Plasmid Preparation, Expressing

    Assessment of Renal Expression of Calcium Regulatory Genes and Proteins. Renal expression of ( A ) Trpv5 , ( B ) Trpv6 and ( C ) Cyp24a1 in wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice (n = 6/group) were assessed by quantitative real-time PCR. All data were normalised to levels of the housekeeping gene Gapdh and wild-type values are expressed as 1. Histogram data are presented as mean ± SEM. # p

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Assessment of Renal Expression of Calcium Regulatory Genes and Proteins. Renal expression of ( A ) Trpv5 , ( B ) Trpv6 and ( C ) Cyp24a1 in wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice (n = 6/group) were assessed by quantitative real-time PCR. All data were normalised to levels of the housekeeping gene Gapdh and wild-type values are expressed as 1. Histogram data are presented as mean ± SEM. # p

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Histological and immunohistochemical assessment of kidneys from HCALC1 mice. Representative images in Trpv5 +/+ (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice of: ( A ) Masson's trichrome staining of renal cortex showing areas of interstitial fibrosis in Trpv5 682P/+ and Trpv5 682P/682P mice (light blue), ( B ) anti-CD3-labelling (green) showing a large number of T-lymphocytes present in the interstitial regions of the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys, ( C ) TUNEL-labelling (green) of the renal cortex showing the presence of tubular cell apoptosis in the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys. Scale bar = 50 µm. ( D ) Immunohistochemical images of kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and NCC (red). * denotes co-localisation. Scale bar = 50 µm. ( E ) Kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and AQP2 (red). * denotes co-localisation. Scale bar = 50 µm.

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Histological and immunohistochemical assessment of kidneys from HCALC1 mice. Representative images in Trpv5 +/+ (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice of: ( A ) Masson's trichrome staining of renal cortex showing areas of interstitial fibrosis in Trpv5 682P/+ and Trpv5 682P/682P mice (light blue), ( B ) anti-CD3-labelling (green) showing a large number of T-lymphocytes present in the interstitial regions of the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys, ( C ) TUNEL-labelling (green) of the renal cortex showing the presence of tubular cell apoptosis in the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys. Scale bar = 50 µm. ( D ) Immunohistochemical images of kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and NCC (red). * denotes co-localisation. Scale bar = 50 µm. ( E ) Kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and AQP2 (red). * denotes co-localisation. Scale bar = 50 µm.

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Immunohistochemistry, Mouse Assay, Staining, TUNEL Assay

    Hypercalciuria in HCALC1 ENU mutant mice and identification of a Trpv5 mutation. ( A ) Urine calcium/creatinine ratios in 23 G2 offspring of the HCALC1 founder male revealed that 10 of the 23 mice were hypercalciuric, consistent with an autosomal dominant inheritance. Bar, mean calcium/creatinine values. ( B ) Haplotype analysis of 89 G2 mice (39 hypercalciuric and 50 normocalciuric) was initially undertaken separately in the hypercalciuric and normocalciuric mice, as the penetrance of HCALC1 was unknown. Haplotype analysis of the hypercalciuric mice localised Hcalc1 to a 17.38 Mb interval on chromosome 6, flanked by rs13478688 and rs30110406 (broken double-headed arrow). Haplotype analysis using combined data for the hypercalciuric and normocalciuric mice identified the smaller interval, 11.94-Mb, flanked by rs13478709 and rs30110406 (solid double-headed arrow). The Hcalc1 locus is inherited with the C57BL/6J haplotype from the F1 founder male. Filled box, C57BL/6J allele; and open box, C3H/HeH allele. Number of mice observed for each haplotype is shown beneath each column. ( C ) DNA sequence analysis of Trpv5 identified a heterozygous T to C transition in codon 682 in hypercalciuric mice predicted to alter a wild-type serine (Ser) to a mutant proline (Pro). This mutation resulted in gain of a Bsa JI restriction enzyme site that was used to confirm the presence of the mutation in the 39 hypercalciuric mice (n = 3 shown) and its absence in the 50 normocalciuric mice (n = 3 shown). wt, wild-type; m, mutant. ( D ) Amino acid sequence alignment revealed evolutionary conservation of the wild-type mouse TRPV5 serine (S) residue at codon 682 (arrowed) in 5 species, as well as in mouse TRPV6 (mTrpv6). Identical residues are shaded black and conservative changes are shaded grey.

    Journal: PLoS ONE

    Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5

    doi: 10.1371/journal.pone.0055412

    Figure Lengend Snippet: Hypercalciuria in HCALC1 ENU mutant mice and identification of a Trpv5 mutation. ( A ) Urine calcium/creatinine ratios in 23 G2 offspring of the HCALC1 founder male revealed that 10 of the 23 mice were hypercalciuric, consistent with an autosomal dominant inheritance. Bar, mean calcium/creatinine values. ( B ) Haplotype analysis of 89 G2 mice (39 hypercalciuric and 50 normocalciuric) was initially undertaken separately in the hypercalciuric and normocalciuric mice, as the penetrance of HCALC1 was unknown. Haplotype analysis of the hypercalciuric mice localised Hcalc1 to a 17.38 Mb interval on chromosome 6, flanked by rs13478688 and rs30110406 (broken double-headed arrow). Haplotype analysis using combined data for the hypercalciuric and normocalciuric mice identified the smaller interval, 11.94-Mb, flanked by rs13478709 and rs30110406 (solid double-headed arrow). The Hcalc1 locus is inherited with the C57BL/6J haplotype from the F1 founder male. Filled box, C57BL/6J allele; and open box, C3H/HeH allele. Number of mice observed for each haplotype is shown beneath each column. ( C ) DNA sequence analysis of Trpv5 identified a heterozygous T to C transition in codon 682 in hypercalciuric mice predicted to alter a wild-type serine (Ser) to a mutant proline (Pro). This mutation resulted in gain of a Bsa JI restriction enzyme site that was used to confirm the presence of the mutation in the 39 hypercalciuric mice (n = 3 shown) and its absence in the 50 normocalciuric mice (n = 3 shown). wt, wild-type; m, mutant. ( D ) Amino acid sequence alignment revealed evolutionary conservation of the wild-type mouse TRPV5 serine (S) residue at codon 682 (arrowed) in 5 species, as well as in mouse TRPV6 (mTrpv6). Identical residues are shaded black and conservative changes are shaded grey.

    Article Snippet: For TRPV5 immuno-detection, 8- µm kidney cryosections were processed for immunofluorescence labelling as previously described ._ENREF_41 Kidney cryosections were co-stained with rabbit anti-TRPV5 (ACC-035, Alomone Labs, Jerusalem, Israel) and goat anti-AQP2 (sc-9882, Santa Cruz, Insight Biotechnology, Wembley, UK) polyclonal antibodies, or with goat anti-TRPV5 (sc-23379, Santa Cruz) and rabbit anti-NCC polyclonal antibodies, followed by the appropriate Alexa Fluor 488- or 594-conjugated secondary antibodies (Molecular Probes).

    Techniques: Mutagenesis, Mouse Assay, Sequencing

    Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Journal: bioRxiv

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

    doi: 10.1101/2020.02.10.941732

    Figure Lengend Snippet: Microscopic images showing localization of TRPV5 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV5 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV5 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV5 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing

    Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Journal: bioRxiv

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

    doi: 10.1101/2020.02.10.941732

    Figure Lengend Snippet: Microscopic images showing localization of TRPV6 in duck sperm. (A-B) Confocal microscopic images depicting the localization of TRPV6 (green) as detected by two different antibodies and Nucleus (blue) by DAPI. Mitochondria (red) is labelled by Mitotracker Red dye in A and C to highlight the channel expression in the mitochondrial region. ( C ) SR-SIM images of TRPV6 localization (using Ab1 antibody) at the head (left) and tail (right) of duck sperm is shown. ( D ) Zoomed up image of neck region of sperm depicting the absence of TRPV6 (green) in the neck region. The head (blue) and arrows mark the start and end point of mitochondrial region.

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.02.10.941732

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

    Article Snippet: Confocal microscopy using two different antibodies against the C-terminus of TRPV5 we found that TRPV5 is exclusively present at the tail (indicated by white arrows), with highest density immediately after the mitochondrial region and intensity gradually decreasing towards the tapering end of the tail ( ).

    Techniques: Expressing, Western Blot, Blocking Assay

    Localisation of TRPV6 protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Localisation of TRPV6 protein in the placentomes and fetal membranes in cows with (RFM) (C, D) and without retained fetal membranes (DFM) (A, B). Strongest labeling of TRPV6 protein was seen in the fetal membranes (FM) of all animals and in the maternal crypt epithelium (ME) of cows with RFM (D).

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Labeling

    Uterine and placentomal TRPV6 and CaBP-9k mRNA expression. TRPV6 and CaBP-9k mRNA expression in the intercaruncular wall (Figure 1A, C) and in the placenta (n ≥ 3 animals/month) (Figure 1B, D) from the 2 nd to 9 th months of pregnancy, determined by real-time PCR. Bars with ** differ significantly (p

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Uterine and placentomal TRPV6 and CaBP-9k mRNA expression. TRPV6 and CaBP-9k mRNA expression in the intercaruncular wall (Figure 1A, C) and in the placenta (n ≥ 3 animals/month) (Figure 1B, D) from the 2 nd to 9 th months of pregnancy, determined by real-time PCR. Bars with ** differ significantly (p

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Western blot analyses of placental TRPV6 and CaBP-9k postpartum in cows with (RFM) and without retained fetal membranes (DFM). A) Representative Western blot of TRPV6 in the placenta. Bands of approximately 75 kDa were detected in all animals. B) Relative expression of TRPV6 was determined by measuring the optical density relative to GAPDH using ImageJ software. Significant changes in expression levels were not detected (p > 0.05). C) Representative Western blot of CaBP-9k. D) The relative expression levels to GAPDH were determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Western blot analyses of placental TRPV6 and CaBP-9k postpartum in cows with (RFM) and without retained fetal membranes (DFM). A) Representative Western blot of TRPV6 in the placenta. Bands of approximately 75 kDa were detected in all animals. B) Relative expression of TRPV6 was determined by measuring the optical density relative to GAPDH using ImageJ software. Significant changes in expression levels were not detected (p > 0.05). C) Representative Western blot of CaBP-9k. D) The relative expression levels to GAPDH were determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Western Blot, Expressing, Software, One-tailed Test

    Immunolabelling of CaBP-9k in placentomes and adherent fetal membranes. Localisation of CaBP-9k during pregnancy with strong signals in the inner layer (IL) of the place tome and in the fetal membranes (FM) compared to the outer labyrinth layer (OL) (A, C). Strong staining in the binucleate trophoblast cells (BNC) and in the maternal epithelium (ME) inside the place tome (B, C). Figure 2 F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Immunolabelling of CaBP-9k in placentomes and adherent fetal membranes. Localisation of CaBP-9k during pregnancy with strong signals in the inner layer (IL) of the place tome and in the fetal membranes (FM) compared to the outer labyrinth layer (OL) (A, C). Strong staining in the binucleate trophoblast cells (BNC) and in the maternal epithelium (ME) inside the place tome (B, C). Figure 2 F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Staining, Negative Control

    Immunolabelling of TRPV6 in the placentomes and adherent fetal membranes. Localisation of TRPV6 during pregnancy in the placenta and adherent membranes with strong staining in the inner layer (IL) of the place tome (A, D) and the adherent fetal membranes (FM) (A, C). Considerably weaker staining in the middle (ML) and outer labyrinth (OL) layers (A, B). Scale bars indicate 200 μm.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Immunolabelling of TRPV6 in the placentomes and adherent fetal membranes. Localisation of TRPV6 during pregnancy in the placenta and adherent membranes with strong staining in the inner layer (IL) of the place tome (A, D) and the adherent fetal membranes (FM) (A, C). Considerably weaker staining in the middle (ML) and outer labyrinth (OL) layers (A, B). Scale bars indicate 200 μm.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Staining

    Immunolabelling of TRPV6 in the uterine endometrium. Localisation of uterine TRPV6 during pregnancy with strong labelling in the glandular (GE) and luminal epithelia (LE). Figure 2F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Immunolabelling of TRPV6 in the uterine endometrium. Localisation of uterine TRPV6 during pregnancy with strong labelling in the glandular (GE) and luminal epithelia (LE). Figure 2F shows a negative control, without anti-TRPV6 treatment. Scale bars indicate 200 μm.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Negative Control

    Placental TRPV6 and CaBP-9k mRNA expression post partum in cows with and without retained fetal membranes. Detection of TRPV6 (A) and Calbindin-D9k (CaBP-9k) (B) mRNA levels in the placenta of postpartum cows that retained the fetal membranes for more than 12 hours (RFM; n = 5) and cows that discharged the fetal membranes (DFM; n = 6), measured by real-time PCR (Taqman). No significant changes were observed (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Placental TRPV6 and CaBP-9k mRNA expression post partum in cows with and without retained fetal membranes. Detection of TRPV6 (A) and Calbindin-D9k (CaBP-9k) (B) mRNA levels in the placenta of postpartum cows that retained the fetal membranes for more than 12 hours (RFM; n = 5) and cows that discharged the fetal membranes (DFM; n = 6), measured by real-time PCR (Taqman). No significant changes were observed (p > 0.05). Data were analyzed by an unpaired one-tailed t-test and are presented as means ± SD.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, One-tailed Test

    Western blot analyses of endometrial and placentomal TRPV6 protein during pregnancy. A) Representative Western blot of TRPV6 protein expression in the uterine wall (n = 3 animals/month). Bands of approximately 75 kDa were detected. An additional band of approximately 90 kDa was seen in the 2 nd and 3 rd months. After incubation with N-glycosidase F (PNGase F), only the smaller band of 75 kDa was seen, demonstrating that the other band is the glycosylated variant of TRPV6. As a negative control, samples were preincubated with a control antigen. B) Relative expression of TRPV6 to GAPDH was determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). C) Representative Western blot analyses of TRPV6 in the placenta from 2 nd to 9 th months of pregnancy (n = 3 animals/month). D) The expression of TRPV6 relative to GAPDH was determined by measuring the optical density using ImageJ software. No significant changes (p > 0.05) were detected. Data were analysed using a parametric one-way analysis of variance (ANOVA), followed by Dunnett’s comparison test, where the second month was taken as the control group, and are presented as means ± SD.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

    doi: 10.1186/1477-7827-10-66

    Figure Lengend Snippet: Western blot analyses of endometrial and placentomal TRPV6 protein during pregnancy. A) Representative Western blot of TRPV6 protein expression in the uterine wall (n = 3 animals/month). Bands of approximately 75 kDa were detected. An additional band of approximately 90 kDa was seen in the 2 nd and 3 rd months. After incubation with N-glycosidase F (PNGase F), only the smaller band of 75 kDa was seen, demonstrating that the other band is the glycosylated variant of TRPV6. As a negative control, samples were preincubated with a control antigen. B) Relative expression of TRPV6 to GAPDH was determined by densitometry using ImageJ software. No significant changes were detected (p > 0.05). C) Representative Western blot analyses of TRPV6 in the placenta from 2 nd to 9 th months of pregnancy (n = 3 animals/month). D) The expression of TRPV6 relative to GAPDH was determined by measuring the optical density using ImageJ software. No significant changes (p > 0.05) were detected. Data were analysed using a parametric one-way analysis of variance (ANOVA), followed by Dunnett’s comparison test, where the second month was taken as the control group, and are presented as means ± SD.

    Article Snippet: As a negative control for the TRPV6 antibody, samples were preincubated with a specific control antigen (Alomone labs, Israel).

    Techniques: Western Blot, Expressing, Incubation, Variant Assay, Negative Control, Software

    Effects of TRPV6 siRNA on the cell motility rate of LNCaP cells. Representative images (A) and graph (B) of the scratch wound-healing assays show that knockdown of TRPV6 suppressed cell motility in LNCaP cells. Cells (1×10 5 ) were seeded into 6-well plates, incubated for 48 hours, transfected with siRNA for TRPV6, and 24 hours later, a constant sized artificial gap was created with a pipette tip in the cell monolayer that was more than 90% saturated. Afterwards, the degree of movement of the cells filling the gap was compared by photographing at 0, 24, and 48 hours with a microscope (magnification ×100). Cells were treated with 1 nM or 100 nM DHT as described in the “MATERIALS AND METHODS” section. DHT, dihydrotestosterone; siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. * p

    Journal: Investigative and Clinical Urology

    Article Title: RNA interference mediated suppression of TRPV6 inhibits the progression of prostate cancer in vitro by modulating cathepsin B and MMP9 expression

    doi: 10.4111/icu.20200511

    Figure Lengend Snippet: Effects of TRPV6 siRNA on the cell motility rate of LNCaP cells. Representative images (A) and graph (B) of the scratch wound-healing assays show that knockdown of TRPV6 suppressed cell motility in LNCaP cells. Cells (1×10 5 ) were seeded into 6-well plates, incubated for 48 hours, transfected with siRNA for TRPV6, and 24 hours later, a constant sized artificial gap was created with a pipette tip in the cell monolayer that was more than 90% saturated. Afterwards, the degree of movement of the cells filling the gap was compared by photographing at 0, 24, and 48 hours with a microscope (magnification ×100). Cells were treated with 1 nM or 100 nM DHT as described in the “MATERIALS AND METHODS” section. DHT, dihydrotestosterone; siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. * p

    Article Snippet: The primary antibodies against TRPV6 used in this experiment were purchased from Alomon Labs (Jerusalem, Israel), whereas the others were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques: Incubation, Transfection, Transferring, Microscopy, Small Interfering RNA

    Effects of TRPV6 siRNA on the proliferation of LNCaP cells. The viability of LNCaP cells was measured by WST-1 assay at 48 hours after transfection with TRPV6 siRNA. Cells were treated with 1 nM or 100 nM DHT as described in the “MATERIALS AND METHODS” section. Cell growth is expressed relative to the value of the negative control, which was set to 100%. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA; DHT, dihydrotestosterone. Data are presented as the mean±standard error of the mean of three independent experiments. # p

    Journal: Investigative and Clinical Urology

    Article Title: RNA interference mediated suppression of TRPV6 inhibits the progression of prostate cancer in vitro by modulating cathepsin B and MMP9 expression

    doi: 10.4111/icu.20200511

    Figure Lengend Snippet: Effects of TRPV6 siRNA on the proliferation of LNCaP cells. The viability of LNCaP cells was measured by WST-1 assay at 48 hours after transfection with TRPV6 siRNA. Cells were treated with 1 nM or 100 nM DHT as described in the “MATERIALS AND METHODS” section. Cell growth is expressed relative to the value of the negative control, which was set to 100%. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA; DHT, dihydrotestosterone. Data are presented as the mean±standard error of the mean of three independent experiments. # p

    Article Snippet: The primary antibodies against TRPV6 used in this experiment were purchased from Alomon Labs (Jerusalem, Israel), whereas the others were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques: WST-1 Assay, Transfection, Negative Control, Small Interfering RNA

    Knockdown efficiency of siTRPV6 in LNCaP cells. (A) The efficacies of three different kinds of small interfering RNA against TRPV6 (siTRPV6#1, siTRPV6#2, and siTRPV6#3) were analyzed by RT-PCR in LNCaP cells. siTRPV6#2 was selected for the following experiments because it blocked more TRPV6 compared with the other variants. The expression of TRPV6 is normalized to β-actin gene expression. (B) The degrees of inhibition of TRPV6 protein levels were analyzed by western blot in siTRPV6#2 transfected LNCaP cells treated with 1 nM or 100 nM DHT. The expression of TRPV6 is normalized to β-actin expression. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; Blank, mock; CON, control scrambled siRNA; DHT, dihydrotestosterone. The data are presented as the mean±standard error of the mean of three independent experiments. * p

    Journal: Investigative and Clinical Urology

    Article Title: RNA interference mediated suppression of TRPV6 inhibits the progression of prostate cancer in vitro by modulating cathepsin B and MMP9 expression

    doi: 10.4111/icu.20200511

    Figure Lengend Snippet: Knockdown efficiency of siTRPV6 in LNCaP cells. (A) The efficacies of three different kinds of small interfering RNA against TRPV6 (siTRPV6#1, siTRPV6#2, and siTRPV6#3) were analyzed by RT-PCR in LNCaP cells. siTRPV6#2 was selected for the following experiments because it blocked more TRPV6 compared with the other variants. The expression of TRPV6 is normalized to β-actin gene expression. (B) The degrees of inhibition of TRPV6 protein levels were analyzed by western blot in siTRPV6#2 transfected LNCaP cells treated with 1 nM or 100 nM DHT. The expression of TRPV6 is normalized to β-actin expression. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; Blank, mock; CON, control scrambled siRNA; DHT, dihydrotestosterone. The data are presented as the mean±standard error of the mean of three independent experiments. * p

    Article Snippet: The primary antibodies against TRPV6 used in this experiment were purchased from Alomon Labs (Jerusalem, Israel), whereas the others were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques: Small Interfering RNA, Reverse Transcription Polymerase Chain Reaction, Expressing, Inhibition, Western Blot, Transfection

    Effect of TRPV6 siRNA on the migratory capability of LNCaP cells. Representative images (A) and graph (B) of the Transwell migration assay of LNCaP cells with TRPV6 siRNA, showing suppressed migration, compared with the scrambled siRNA control. Cells were treated with 1 nM DHT as described in the “MATERIALS AND METHODS” section. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. * p

    Journal: Investigative and Clinical Urology

    Article Title: RNA interference mediated suppression of TRPV6 inhibits the progression of prostate cancer in vitro by modulating cathepsin B and MMP9 expression

    doi: 10.4111/icu.20200511

    Figure Lengend Snippet: Effect of TRPV6 siRNA on the migratory capability of LNCaP cells. Representative images (A) and graph (B) of the Transwell migration assay of LNCaP cells with TRPV6 siRNA, showing suppressed migration, compared with the scrambled siRNA control. Cells were treated with 1 nM DHT as described in the “MATERIALS AND METHODS” section. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. * p

    Article Snippet: The primary antibodies against TRPV6 used in this experiment were purchased from Alomon Labs (Jerusalem, Israel), whereas the others were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques: Transwell Migration Assay, Migration, Small Interfering RNA

    Effect of TRPV6 siRNA on the invasive capability of LNCaP cells. Representative images (A) and graph (B) of the Transwell invasion of LNCaP cells with TRPV6 inhibition and respective scrambled siRNA control. (C) Effect of TRPV6 siRNA on the protein expression of cathepsin B matrix metalloproteinase 9 (MMP9) in LNCaP cells. Both cathepsin B and MMP9 expression were reduced by TRPV6 siRNA transfection in LNCaP cells. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; Blank, mock; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. # p

    Journal: Investigative and Clinical Urology

    Article Title: RNA interference mediated suppression of TRPV6 inhibits the progression of prostate cancer in vitro by modulating cathepsin B and MMP9 expression

    doi: 10.4111/icu.20200511

    Figure Lengend Snippet: Effect of TRPV6 siRNA on the invasive capability of LNCaP cells. Representative images (A) and graph (B) of the Transwell invasion of LNCaP cells with TRPV6 inhibition and respective scrambled siRNA control. (C) Effect of TRPV6 siRNA on the protein expression of cathepsin B matrix metalloproteinase 9 (MMP9) in LNCaP cells. Both cathepsin B and MMP9 expression were reduced by TRPV6 siRNA transfection in LNCaP cells. siTRPV6, small interfering RNA targeting transient receptor potential cation channel vanilloid subfamily number 6; Blank, mock; CON, control scrambled siRNA. Data are presented as the mean±standard error of the mean of three independent experiments. # p

    Article Snippet: The primary antibodies against TRPV6 used in this experiment were purchased from Alomon Labs (Jerusalem, Israel), whereas the others were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques: Inhibition, Expressing, Transfection, Small Interfering RNA