rabbit anti aqp6 polyclonal antibody Search Results


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  • 80
    Alpha Diagnostics anti aqp6 rabbit polyclonal igg
    Hydrogen peroxide transport in HeLa cells with <t>AQP6</t> reduced expression in the presence and in the absence of CNPs treatment. ( A ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into mock-transfected HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( B ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into AQP6-silenced (siRNA AQP6) HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( C ) Time course of H 2 O 2 fluorescence into mock- and siRNA-transfected HeLa cells, in the absence (Ctr) and in the presence of CNPs treatment (CNPs). Curves start when 50 μM H 2 O 2 was added. Results are the mean of at least 3 different experiments and SEMs were omitted for clarity. ( D ) Maximal H 2 O 2 fluorescence values were obtained by computerized least squares regression, fitting the experimental points of the time courses of H 2 O 2 transported curves with a one-phase exponential association equation (GraphPad Prism 4.00, 2003). a , p
    Anti Aqp6 Rabbit Polyclonal Igg, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti aqp6 rabbit polyclonal igg/product/Alpha Diagnostics
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    anti aqp6 rabbit polyclonal igg - by Bioz Stars, 2022-12
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    80
    Alpha Diagnostics affinity pure anti aqp6 rabbit polyclonal primary antibody
    Representative images of confocal laser scanning microscopy and 3D colocalization analysis of <t>AQP6</t> ( A–D ) with concanavalin A (ConA) in MeT-5A, REN, and MSTO-211H cell lines. Green labeling indicates the presence of ConA ( B ), red labeling indicates the expression of AQP6 ( C ), while DAPI (blue, D ) indicates counterstained nuclei. Yellow labelling shows the colocalization signal of AQP with ConA ( A ). Scale bar: 10 μm (right panel). Statistical analysis of Pearson’s correlation coefficient r, Manders’ colocalization coefficient (M1 and M2), and Van Steensel’s maxima cross-correlation function (CCF) were obtained from 4 different double immunofluorescence experiments with <t>anti-AQP6</t> antibody and anti-AQP6 antibodies. Coefficients were determined by 3D analysis of at least 20 cells for each cell line (8–15 z-stack for image) using the JACoP plugin of Fiji. The columns represent the mean ± SD of the coefficient values; *, p
    Affinity Pure Anti Aqp6 Rabbit Polyclonal Primary Antibody, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/affinity pure anti aqp6 rabbit polyclonal primary antibody/product/Alpha Diagnostics
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    affinity pure anti aqp6 rabbit polyclonal primary antibody - by Bioz Stars, 2022-12
    80/100 stars
      Buy from Supplier

    85
    Alomone Labs rabbit anti aqp6 polyclonal antibody
    Characterization of M-1 cellular expression of aquaporin A) Characterization of <t>AQP6</t> mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. An AQP6 specific band (arrowhead) was amplified from M-1 cellular total RNA (lane 1), M-1 cellular genomic DNA (gDNA, lane 3) and normal mouse kidney total RNA (MK cDNA, lane 4). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot demonstrating the AQP6 specific band (arrowhead) and glycosylated band (double arrowhead) in total membrane protein extracted from M-1 cells (lane 1) and murine kidney (MK, lane 2). B) Characterization of AQP2 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. AQP2 mRNA was not amplified from M-1 cellular total RNA (lane 1), although it was observed in M-1 gDNA (lane 3) and MK cDNA (lane 4). Similarly, AQP2 protein was not observed in total membrane protein isolated from M-1 cells (lane 1), although it was observed in total membrane protein isolated from the MK (lane 2). An arrowhead and a double arrow indicate the unglycosylated and glycosylated bands, respectively. Immunofluorescence showed the M-1 cells were positive to the AQP6 antibody (A, lower column), but negative to AQP2 antibody (B, lower column). Bar=50 µ m.
    Rabbit Anti Aqp6 Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti aqp6 polyclonal antibody/product/Alomone Labs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti aqp6 polyclonal antibody - by Bioz Stars, 2022-12
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    93
    Alomone Labs rabbit anti aqp2
    Characterization of M-1 cellular expression of aquaporin A) Characterization of <t>AQP6</t> mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. An AQP6 specific band (arrowhead) was amplified from M-1 cellular total RNA (lane 1), M-1 cellular genomic DNA (gDNA, lane 3) and normal mouse kidney total RNA (MK cDNA, lane 4). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot demonstrating the AQP6 specific band (arrowhead) and glycosylated band (double arrowhead) in total membrane protein extracted from M-1 cells (lane 1) and murine kidney (MK, lane 2). B) Characterization of AQP2 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. AQP2 mRNA was not amplified from M-1 cellular total RNA (lane 1), although it was observed in M-1 gDNA (lane 3) and MK cDNA (lane 4). Similarly, AQP2 protein was not observed in total membrane protein isolated from M-1 cells (lane 1), although it was observed in total membrane protein isolated from the MK (lane 2). An arrowhead and a double arrow indicate the unglycosylated and glycosylated bands, respectively. Immunofluorescence showed the M-1 cells were positive to the AQP6 antibody (A, lower column), but negative to AQP2 antibody (B, lower column). Bar=50 µ m.
    Rabbit Anti Aqp2, 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
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    Image Search Results


    Hydrogen peroxide transport in HeLa cells with AQP6 reduced expression in the presence and in the absence of CNPs treatment. ( A ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into mock-transfected HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( B ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into AQP6-silenced (siRNA AQP6) HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( C ) Time course of H 2 O 2 fluorescence into mock- and siRNA-transfected HeLa cells, in the absence (Ctr) and in the presence of CNPs treatment (CNPs). Curves start when 50 μM H 2 O 2 was added. Results are the mean of at least 3 different experiments and SEMs were omitted for clarity. ( D ) Maximal H 2 O 2 fluorescence values were obtained by computerized least squares regression, fitting the experimental points of the time courses of H 2 O 2 transported curves with a one-phase exponential association equation (GraphPad Prism 4.00, 2003). a , p

    Journal: International Journal of Molecular Sciences

    Article Title: Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability

    doi: 10.3390/ijms231810837

    Figure Lengend Snippet: Hydrogen peroxide transport in HeLa cells with AQP6 reduced expression in the presence and in the absence of CNPs treatment. ( A ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into mock-transfected HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( B ) Representative frames extracted from videos showing the kinetics of H 2 O 2 transport into AQP6-silenced (siRNA AQP6) HeLa cells before (left panel) and after (right panel) addition of 50 μM H 2 O 2 in the absence (Ctr) and in the presence of CNPs treatment (CNPs). The increased HyPer7-NES fluorescence is shown in pseudocolor (upper panel, the scale used is indicated in the insert). ( C ) Time course of H 2 O 2 fluorescence into mock- and siRNA-transfected HeLa cells, in the absence (Ctr) and in the presence of CNPs treatment (CNPs). Curves start when 50 μM H 2 O 2 was added. Results are the mean of at least 3 different experiments and SEMs were omitted for clarity. ( D ) Maximal H 2 O 2 fluorescence values were obtained by computerized least squares regression, fitting the experimental points of the time courses of H 2 O 2 transported curves with a one-phase exponential association equation (GraphPad Prism 4.00, 2003). a , p

    Article Snippet: After permeabilization by incubating at 80 °C for 30 min with the retrieval buffer (10 mM citrate-HCl, pH 6.0), the samples were blocked with 3% BSA in PBS at RT for 30 min. Coverslips were incubated overnight at 4 °C with the affinity pure anti-AQP3 rabbit antibody (ab125045, 1:400 dilution; Abcam, Cambridge, UK), anti-AQP6 rabbit polyclonal IgG (AQP61-A, 1:200 dilution; Alpha Diagnostic International, San Antonio, TX, USA), anti-AQP8 rabbit antibody (HPA046259, 1:500 dilution; Sigma-Aldrich, St. Louis, MO, U.S.A.), and anti-AQP11 rabbit antibody (ab122821, 1:50 dilution; Abcam, Cambridge, UK).

    Techniques: Expressing, Transfection, Fluorescence

    Expression of AQP3, AQP6, AQP8, and AQP11 proteins in HeLa cells. Blots representative of three were shown. Each lane was loaded with 30 μg of proteins and probed with affinity-purified antibodies as described in the Materials and Methods. The same blots were stripped and reprobed with an anti-β-2-microglobulin (β2M) antibody, as housekeeping. Major bands of the expected molecular weights are shown.

    Journal: International Journal of Molecular Sciences

    Article Title: Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability

    doi: 10.3390/ijms231810837

    Figure Lengend Snippet: Expression of AQP3, AQP6, AQP8, and AQP11 proteins in HeLa cells. Blots representative of three were shown. Each lane was loaded with 30 μg of proteins and probed with affinity-purified antibodies as described in the Materials and Methods. The same blots were stripped and reprobed with an anti-β-2-microglobulin (β2M) antibody, as housekeeping. Major bands of the expected molecular weights are shown.

    Article Snippet: After permeabilization by incubating at 80 °C for 30 min with the retrieval buffer (10 mM citrate-HCl, pH 6.0), the samples were blocked with 3% BSA in PBS at RT for 30 min. Coverslips were incubated overnight at 4 °C with the affinity pure anti-AQP3 rabbit antibody (ab125045, 1:400 dilution; Abcam, Cambridge, UK), anti-AQP6 rabbit polyclonal IgG (AQP61-A, 1:200 dilution; Alpha Diagnostic International, San Antonio, TX, USA), anti-AQP8 rabbit antibody (HPA046259, 1:500 dilution; Sigma-Aldrich, St. Louis, MO, U.S.A.), and anti-AQP11 rabbit antibody (ab122821, 1:50 dilution; Abcam, Cambridge, UK).

    Techniques: Expressing, Affinity Purification

    Representative images of confocal laser scanning microscopy of AQP3, AQP6, AQP8, and AQP11 with CNPs in HeLa cells. Green labeling indicates the presence of AQPs ( A ), red labeling the CNPs ( B ), and DAPI (blue; C ) counterstained nuclei. Yellow labeling shows the colocalization signal of AQP with CNPs ( D ). Scale bar, 10 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability

    doi: 10.3390/ijms231810837

    Figure Lengend Snippet: Representative images of confocal laser scanning microscopy of AQP3, AQP6, AQP8, and AQP11 with CNPs in HeLa cells. Green labeling indicates the presence of AQPs ( A ), red labeling the CNPs ( B ), and DAPI (blue; C ) counterstained nuclei. Yellow labeling shows the colocalization signal of AQP with CNPs ( D ). Scale bar, 10 μm.

    Article Snippet: After permeabilization by incubating at 80 °C for 30 min with the retrieval buffer (10 mM citrate-HCl, pH 6.0), the samples were blocked with 3% BSA in PBS at RT for 30 min. Coverslips were incubated overnight at 4 °C with the affinity pure anti-AQP3 rabbit antibody (ab125045, 1:400 dilution; Abcam, Cambridge, UK), anti-AQP6 rabbit polyclonal IgG (AQP61-A, 1:200 dilution; Alpha Diagnostic International, San Antonio, TX, USA), anti-AQP8 rabbit antibody (HPA046259, 1:500 dilution; Sigma-Aldrich, St. Louis, MO, U.S.A.), and anti-AQP11 rabbit antibody (ab122821, 1:50 dilution; Abcam, Cambridge, UK).

    Techniques: Confocal Laser Scanning Microscopy, Labeling

    Three-dimensional colocalization analysis of AQP3, AQP6, AQP8, and AQP11 with CNPs in HeLa cells. Statistical analysis of Pearson’s correlation coefficient r ( A ), Van Steensel’s maxima cross-correlation function (CCFmax) ( B ), and Manders’ colocalization coefficients (M1 and M2) ( C , D ) were obtained from 4 different double immunofluorescence experiments with anti-AQP antibodies and red-labeled CNPs (see Materials and Methods). Coefficients were determined by 3D analysis of at least 20 cells for each cell line (8–15 z-stack for image) using the JACoP plugin of Fiji. The columns represent the mean ± SEM of the coefficient values (Brown–Forsythe and Welch ANOVA tests followed by Dunnett T3 post-test). a , p

    Journal: International Journal of Molecular Sciences

    Article Title: Cerium Oxide Nanoparticles Regulate Oxidative Stress in HeLa Cells by Increasing the Aquaporin-Mediated Hydrogen Peroxide Permeability

    doi: 10.3390/ijms231810837

    Figure Lengend Snippet: Three-dimensional colocalization analysis of AQP3, AQP6, AQP8, and AQP11 with CNPs in HeLa cells. Statistical analysis of Pearson’s correlation coefficient r ( A ), Van Steensel’s maxima cross-correlation function (CCFmax) ( B ), and Manders’ colocalization coefficients (M1 and M2) ( C , D ) were obtained from 4 different double immunofluorescence experiments with anti-AQP antibodies and red-labeled CNPs (see Materials and Methods). Coefficients were determined by 3D analysis of at least 20 cells for each cell line (8–15 z-stack for image) using the JACoP plugin of Fiji. The columns represent the mean ± SEM of the coefficient values (Brown–Forsythe and Welch ANOVA tests followed by Dunnett T3 post-test). a , p

    Article Snippet: After permeabilization by incubating at 80 °C for 30 min with the retrieval buffer (10 mM citrate-HCl, pH 6.0), the samples were blocked with 3% BSA in PBS at RT for 30 min. Coverslips were incubated overnight at 4 °C with the affinity pure anti-AQP3 rabbit antibody (ab125045, 1:400 dilution; Abcam, Cambridge, UK), anti-AQP6 rabbit polyclonal IgG (AQP61-A, 1:200 dilution; Alpha Diagnostic International, San Antonio, TX, USA), anti-AQP8 rabbit antibody (HPA046259, 1:500 dilution; Sigma-Aldrich, St. Louis, MO, U.S.A.), and anti-AQP11 rabbit antibody (ab122821, 1:50 dilution; Abcam, Cambridge, UK).

    Techniques: Immunofluorescence, Labeling

    Representative images of confocal laser scanning microscopy and 3D colocalization analysis of AQP6 ( A–D ) with concanavalin A (ConA) in MeT-5A, REN, and MSTO-211H cell lines. Green labeling indicates the presence of ConA ( B ), red labeling indicates the expression of AQP6 ( C ), while DAPI (blue, D ) indicates counterstained nuclei. Yellow labelling shows the colocalization signal of AQP with ConA ( A ). Scale bar: 10 μm (right panel). Statistical analysis of Pearson’s correlation coefficient r, Manders’ colocalization coefficient (M1 and M2), and Van Steensel’s maxima cross-correlation function (CCF) were obtained from 4 different double immunofluorescence experiments with anti-AQP6 antibody and anti-AQP6 antibodies. Coefficients were determined by 3D analysis of at least 20 cells for each cell line (8–15 z-stack for image) using the JACoP plugin of Fiji. The columns represent the mean ± SD of the coefficient values; *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Representative images of confocal laser scanning microscopy and 3D colocalization analysis of AQP6 ( A–D ) with concanavalin A (ConA) in MeT-5A, REN, and MSTO-211H cell lines. Green labeling indicates the presence of ConA ( B ), red labeling indicates the expression of AQP6 ( C ), while DAPI (blue, D ) indicates counterstained nuclei. Yellow labelling shows the colocalization signal of AQP with ConA ( A ). Scale bar: 10 μm (right panel). Statistical analysis of Pearson’s correlation coefficient r, Manders’ colocalization coefficient (M1 and M2), and Van Steensel’s maxima cross-correlation function (CCF) were obtained from 4 different double immunofluorescence experiments with anti-AQP6 antibody and anti-AQP6 antibodies. Coefficients were determined by 3D analysis of at least 20 cells for each cell line (8–15 z-stack for image) using the JACoP plugin of Fiji. The columns represent the mean ± SD of the coefficient values; *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Confocal Laser Scanning Microscopy, Labeling, Expressing, Immunofluorescence

    Effect of oxidative stress on water pe rmeability of MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cell lines after AQP6 gene silencing. Mock-transfected (scrambled, mock-transfected) and silenced cells (siRNA AQP6) were exposed to 150 mOsm osmotic gradients in two different conditions: untreated cells (Ctr) and cells treated with H 2 O 2 (H 2 O 2 ). *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Effect of oxidative stress on water pe rmeability of MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cell lines after AQP6 gene silencing. Mock-transfected (scrambled, mock-transfected) and silenced cells (siRNA AQP6) were exposed to 150 mOsm osmotic gradients in two different conditions: untreated cells (Ctr) and cells treated with H 2 O 2 (H 2 O 2 ). *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Transfection

    qRT-PCR reaction analysis of AQP3, AQP5, AQP6, AQP8, AQP9, and AQP11 expression in mesothelial (MeT-5A, in white), epithelioid, and biphasic MPM cell lines (REN and MSTO-211H, in blue and green, respectively). Bars represent the mean ± SEM of fold change values (n = 4). *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: qRT-PCR reaction analysis of AQP3, AQP5, AQP6, AQP8, AQP9, and AQP11 expression in mesothelial (MeT-5A, in white), epithelioid, and biphasic MPM cell lines (REN and MSTO-211H, in blue and green, respectively). Bars represent the mean ± SEM of fold change values (n = 4). *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Quantitative RT-PCR, Expressing

    Effect of aquaporin-6 (AQP6) silencing on the cell proliferation of MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cultured cells. Cell growth was evaluated by measuring the OD at 570 nm at 24, 48, and 72 h after cell silencing compared with scrambled silenced control cells. The initial cell number was about 25,000 cells/well. AQP6-null cells (siRNA AQP6) showed a significantly decreased proliferation compared with controls after 24 and 48 h from silencing in Met-5A and MSTO-211H cells and after 24, 48, and 72 h from silencing in REN cells. Values (OD at 570 nm normalized to the cell number) are mean ± SEM of cells for each of 4 different experiments. *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Effect of aquaporin-6 (AQP6) silencing on the cell proliferation of MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cultured cells. Cell growth was evaluated by measuring the OD at 570 nm at 24, 48, and 72 h after cell silencing compared with scrambled silenced control cells. The initial cell number was about 25,000 cells/well. AQP6-null cells (siRNA AQP6) showed a significantly decreased proliferation compared with controls after 24 and 48 h from silencing in Met-5A and MSTO-211H cells and after 24, 48, and 72 h from silencing in REN cells. Values (OD at 570 nm normalized to the cell number) are mean ± SEM of cells for each of 4 different experiments. *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Cell Culture

    Effect of oxidative stress on cell proliferation of MeT-5A, REN, and MSTO-211H cells silenced for aquaporin-6 (siRNA) and mock-transfected (Ctr). Cell growth was evaluated by measuring the OD at 570 nm at 48 h after cell silencing compared with mock-transfected control cells. Oxidative stress was produced in siRNA and mock-transfected cells, as indicated in Materials and Methods. Values (expressed as the percent of proliferation) are means ± SEM of cells for each of 4 different experiments. *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Effect of oxidative stress on cell proliferation of MeT-5A, REN, and MSTO-211H cells silenced for aquaporin-6 (siRNA) and mock-transfected (Ctr). Cell growth was evaluated by measuring the OD at 570 nm at 48 h after cell silencing compared with mock-transfected control cells. Oxidative stress was produced in siRNA and mock-transfected cells, as indicated in Materials and Methods. Values (expressed as the percent of proliferation) are means ± SEM of cells for each of 4 different experiments. *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Transfection, Produced

    Effect of aquaporin-6 (AQP6) silencing on the H 2 O 2 permeability MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cell lines. ( A – C ) HeLa cells were silenced with AQP6 siRNA and then transiently transfected with HyPer7 sensor, as described in Materials and Methods. Control (scrambled; CTR) and silenced cells (siRNA) were exposed to 50 μM H 2 O 2 gradient (final concentration). Curves show the time course of H 2 O 2 transported into the cells after H 2 O 2 injection (red arrow). ( D ) Bars represent the H 2 O 2 permeability of cells expressed as a percent of maximal fluorescence. Values are means ± SEM of cells for each of 3 different experiments in triplicates. *, p = 0.0414, p = 0.0260, p = 0.0275 versus Ctr, for Met-5A, REN, and MSTO-211H, respectively (Brown–Forsythe and Welch ANOVA tests).

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Effect of aquaporin-6 (AQP6) silencing on the H 2 O 2 permeability MeT-5A ( A ), REN ( B ), and MSTO-211H ( C ) cell lines. ( A – C ) HeLa cells were silenced with AQP6 siRNA and then transiently transfected with HyPer7 sensor, as described in Materials and Methods. Control (scrambled; CTR) and silenced cells (siRNA) were exposed to 50 μM H 2 O 2 gradient (final concentration). Curves show the time course of H 2 O 2 transported into the cells after H 2 O 2 injection (red arrow). ( D ) Bars represent the H 2 O 2 permeability of cells expressed as a percent of maximal fluorescence. Values are means ± SEM of cells for each of 3 different experiments in triplicates. *, p = 0.0414, p = 0.0260, p = 0.0275 versus Ctr, for Met-5A, REN, and MSTO-211H, respectively (Brown–Forsythe and Welch ANOVA tests).

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Permeability, Transfection, Concentration Assay, Injection, Fluorescence

    Immunoblotting and densitometric analysis of AQP3 ( A ), AQP5 ( B ), AQP6 ( C ), AQP8 ( D ), AQP9 ( E ), and AQP11 ( F ) in mesothelial (MeT-5A, in white), epithelioid, and biphasic MPM cell lines (REN and MSTO-211H, in blue and green, respectively). ( Left panels ) Representative blots of three different experiments are shown. Lanes were loaded with 30 μg of proteins, probed with affinity-purified antibodies, and processed as described in Materials and Methods. The same blots were stripped and re-probed with anti-β-actin (BAC) antibody, as housekeeping. Major bands of the expected molecular weights are shown. ( Right panels ) Densitometry of AQP protein levels in the three cell lines. Each bar represents the mean ± SEM of the normalized values of AQP protein expression. *, p

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Immunoblotting and densitometric analysis of AQP3 ( A ), AQP5 ( B ), AQP6 ( C ), AQP8 ( D ), AQP9 ( E ), and AQP11 ( F ) in mesothelial (MeT-5A, in white), epithelioid, and biphasic MPM cell lines (REN and MSTO-211H, in blue and green, respectively). ( Left panels ) Representative blots of three different experiments are shown. Lanes were loaded with 30 μg of proteins, probed with affinity-purified antibodies, and processed as described in Materials and Methods. The same blots were stripped and re-probed with anti-β-actin (BAC) antibody, as housekeeping. Major bands of the expected molecular weights are shown. ( Right panels ) Densitometry of AQP protein levels in the three cell lines. Each bar represents the mean ± SEM of the normalized values of AQP protein expression. *, p

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Affinity Purification, BAC Assay, Expressing

    Immunocytochemical localization of AQP3 ( A ), AQP5 ( B ), and AQP6 ( C ) proteins in MeT-5A, REN, and MSTO-211H cell lines. AQP3 and AQP5 staining appeared to be confined mainly to intracellular structures. AQP6 staining in MeT-5A is mainly intracellular and has a low expression in the plasma membrane. REN and MSTO-211H cell lines showed strong labelling in discrete areas of the plasma membrane, in addition to the intracellular expression. Arrowheads indicate the localization in plasma membranes. Scale bar: 20 μm.

    Journal: Cells

    Article Title: Aquaporin-6 May Increase the Resistance to Oxidative Stress of Malignant Pleural Mesothelioma Cells

    doi: 10.3390/cells11121892

    Figure Lengend Snippet: Immunocytochemical localization of AQP3 ( A ), AQP5 ( B ), and AQP6 ( C ) proteins in MeT-5A, REN, and MSTO-211H cell lines. AQP3 and AQP5 staining appeared to be confined mainly to intracellular structures. AQP6 staining in MeT-5A is mainly intracellular and has a low expression in the plasma membrane. REN and MSTO-211H cell lines showed strong labelling in discrete areas of the plasma membrane, in addition to the intracellular expression. Arrowheads indicate the localization in plasma membranes. Scale bar: 20 μm.

    Article Snippet: The cells were then blocked with 3% BSA in PBS at room temperature for 30 min. Coverslips were incubated overnight at 4 °C with affinity pure anti-AQP6 rabbit polyclonal primary antibody (# AQP61-A, 1:250; Alpha Diagnostic International, San Antonio, TX, USA).

    Techniques: Staining, Expressing

    Characterization of M-1 cellular expression of aquaporin A) Characterization of AQP6 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. An AQP6 specific band (arrowhead) was amplified from M-1 cellular total RNA (lane 1), M-1 cellular genomic DNA (gDNA, lane 3) and normal mouse kidney total RNA (MK cDNA, lane 4). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot demonstrating the AQP6 specific band (arrowhead) and glycosylated band (double arrowhead) in total membrane protein extracted from M-1 cells (lane 1) and murine kidney (MK, lane 2). B) Characterization of AQP2 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. AQP2 mRNA was not amplified from M-1 cellular total RNA (lane 1), although it was observed in M-1 gDNA (lane 3) and MK cDNA (lane 4). Similarly, AQP2 protein was not observed in total membrane protein isolated from M-1 cells (lane 1), although it was observed in total membrane protein isolated from the MK (lane 2). An arrowhead and a double arrow indicate the unglycosylated and glycosylated bands, respectively. Immunofluorescence showed the M-1 cells were positive to the AQP6 antibody (A, lower column), but negative to AQP2 antibody (B, lower column). Bar=50 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Expression of Adrenergic and Cholinergic Receptors in Murine Renal Intercalated Cells

    doi: 10.1292/jvms.14-0315

    Figure Lengend Snippet: Characterization of M-1 cellular expression of aquaporin A) Characterization of AQP6 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. An AQP6 specific band (arrowhead) was amplified from M-1 cellular total RNA (lane 1), M-1 cellular genomic DNA (gDNA, lane 3) and normal mouse kidney total RNA (MK cDNA, lane 4). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot demonstrating the AQP6 specific band (arrowhead) and glycosylated band (double arrowhead) in total membrane protein extracted from M-1 cells (lane 1) and murine kidney (MK, lane 2). B) Characterization of AQP2 mRNA and protein expression in M-1 cells by RT-PCR (upper column) and immunoblot (middle column), respectively. AQP2 mRNA was not amplified from M-1 cellular total RNA (lane 1), although it was observed in M-1 gDNA (lane 3) and MK cDNA (lane 4). Similarly, AQP2 protein was not observed in total membrane protein isolated from M-1 cells (lane 1), although it was observed in total membrane protein isolated from the MK (lane 2). An arrowhead and a double arrow indicate the unglycosylated and glycosylated bands, respectively. Immunofluorescence showed the M-1 cells were positive to the AQP6 antibody (A, lower column), but negative to AQP2 antibody (B, lower column). Bar=50 µ m.

    Article Snippet: Membranes were incubated with diluted primary antibodies including rabbit anti-AQP6 polyclonal antibody (pAb, #AQP-006, Alomone Labs, Jerusalem, Israel), rabbit anti-AQP2 pAb (#AQP-002, Alomone labs), rabbit anti-ADR b2 pAb (#Sc-569, Santa Cruz, TX, U.S.A.) and rabbit anti-mAChR m1 pAb (#010, Alomone Labs) for 2 hr at room temperature or at 4°C overnight.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Negative Control, Isolation, Immunofluorescence

    Expression of adrenergic receptors in M-1 cells and histological localization of the β2 receptor in the mouse kidney. A) RT-PCR (upper column) of ADRs in M-1 total cellular RNA (cDNA, lane 1) and genomic DNA (gDNA, lane 3). The α2 (second row) and β2 (third row) ADRs were amplified from M-1 total cellular RNA, but the α1 (first row) and β3 (fourth row) ADRs were not. No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot of the β2 receptor (lower column) shows a specific 60–75 kDa band (arrowhead). B) and C) Immunohistochemistry of the β2 receptor (B) and AQP6 (C) in the mouse renal cortex (arrows indicate the same points on both images). Arrows indicate AQP6-positive, β2-positive cells, and arrowheads indicate AQP6-positive, β2-negative cells in the CCDs. D) and E) Immunohistochemistry of the β2 receptor (D) and AQP6 (E) in the outer medulla. β2 receptor immunoreactivities were observed to the straight segment of proximal tubules, but not in the collecting ducts. Arrowheads (AQP6 positive cells) indicate the same points on both images. G: glomerulus. Bar=20 µ m (B and C) and 100 µ m (D and E).

    Journal: The Journal of Veterinary Medical Science

    Article Title: Expression of Adrenergic and Cholinergic Receptors in Murine Renal Intercalated Cells

    doi: 10.1292/jvms.14-0315

    Figure Lengend Snippet: Expression of adrenergic receptors in M-1 cells and histological localization of the β2 receptor in the mouse kidney. A) RT-PCR (upper column) of ADRs in M-1 total cellular RNA (cDNA, lane 1) and genomic DNA (gDNA, lane 3). The α2 (second row) and β2 (third row) ADRs were amplified from M-1 total cellular RNA, but the α1 (first row) and β3 (fourth row) ADRs were not. No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot of the β2 receptor (lower column) shows a specific 60–75 kDa band (arrowhead). B) and C) Immunohistochemistry of the β2 receptor (B) and AQP6 (C) in the mouse renal cortex (arrows indicate the same points on both images). Arrows indicate AQP6-positive, β2-positive cells, and arrowheads indicate AQP6-positive, β2-negative cells in the CCDs. D) and E) Immunohistochemistry of the β2 receptor (D) and AQP6 (E) in the outer medulla. β2 receptor immunoreactivities were observed to the straight segment of proximal tubules, but not in the collecting ducts. Arrowheads (AQP6 positive cells) indicate the same points on both images. G: glomerulus. Bar=20 µ m (B and C) and 100 µ m (D and E).

    Article Snippet: Membranes were incubated with diluted primary antibodies including rabbit anti-AQP6 polyclonal antibody (pAb, #AQP-006, Alomone Labs, Jerusalem, Israel), rabbit anti-AQP2 pAb (#AQP-002, Alomone labs), rabbit anti-ADR b2 pAb (#Sc-569, Santa Cruz, TX, U.S.A.) and rabbit anti-mAChR m1 pAb (#010, Alomone Labs) for 2 hr at room temperature or at 4°C overnight.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Negative Control, Immunohistochemistry

    Expression of mAChRs in M-1 cells and immunohistochemical localization of the m1 ADR in the mouse kidney. A) RT-PCR (upper column) of m1, m3, m4 and m5 receptor subtypes in M-1 total cellular RNA (lane 1, arrowheads). Genomic DNA was used as a control (lane 3). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot (lower column) demonstrating the presence of the m1 receptor subtype in the M-1 total membrane protein fraction. The most intense band is observed at 55 to 70 kDa (arrowhead). B) to E) Immunohistochemistry demonstrating the presence of the m1 mAChR (B and D) and AQP6 (C and E) in the cortex (B and C) and renal outer medulla (D and E). Arrows indicate AQP6-positive, m1-positive cells. Arrowheads indicate AQP6-positive, m1-negative cells. Reactions of m1 antibody were stronger to the medullary collecting ducts (B) than to the CCDs (D). Bar=20 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Expression of Adrenergic and Cholinergic Receptors in Murine Renal Intercalated Cells

    doi: 10.1292/jvms.14-0315

    Figure Lengend Snippet: Expression of mAChRs in M-1 cells and immunohistochemical localization of the m1 ADR in the mouse kidney. A) RT-PCR (upper column) of m1, m3, m4 and m5 receptor subtypes in M-1 total cellular RNA (lane 1, arrowheads). Genomic DNA was used as a control (lane 3). No positive band was observed in RTase (−) negative control lane (lane 2). Immunoblot (lower column) demonstrating the presence of the m1 receptor subtype in the M-1 total membrane protein fraction. The most intense band is observed at 55 to 70 kDa (arrowhead). B) to E) Immunohistochemistry demonstrating the presence of the m1 mAChR (B and D) and AQP6 (C and E) in the cortex (B and C) and renal outer medulla (D and E). Arrows indicate AQP6-positive, m1-positive cells. Arrowheads indicate AQP6-positive, m1-negative cells. Reactions of m1 antibody were stronger to the medullary collecting ducts (B) than to the CCDs (D). Bar=20 µ m.

    Article Snippet: Membranes were incubated with diluted primary antibodies including rabbit anti-AQP6 polyclonal antibody (pAb, #AQP-006, Alomone Labs, Jerusalem, Israel), rabbit anti-AQP2 pAb (#AQP-002, Alomone labs), rabbit anti-ADR b2 pAb (#Sc-569, Santa Cruz, TX, U.S.A.) and rabbit anti-mAChR m1 pAb (#010, Alomone Labs) for 2 hr at room temperature or at 4°C overnight.

    Techniques: Expressing, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Negative Control

    Dual staining of AChE and AQP6 in CCDs. A) AChE was localized to the glomerulus (G) and the nerve fibers along the interlobular arterioles (arrows) B) AChE in CCD cells was stained with the modified Karnovsky-Roots method (arrows). C) AQP6 in the mouse renal cortex was stained by immunohistochemistry (arrowheads). AChE and AQP6 staining did not overlap in the CCDs. The arrows and arrowheads indicate identical cells in panels A and B. Bar=20 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Expression of Adrenergic and Cholinergic Receptors in Murine Renal Intercalated Cells

    doi: 10.1292/jvms.14-0315

    Figure Lengend Snippet: Dual staining of AChE and AQP6 in CCDs. A) AChE was localized to the glomerulus (G) and the nerve fibers along the interlobular arterioles (arrows) B) AChE in CCD cells was stained with the modified Karnovsky-Roots method (arrows). C) AQP6 in the mouse renal cortex was stained by immunohistochemistry (arrowheads). AChE and AQP6 staining did not overlap in the CCDs. The arrows and arrowheads indicate identical cells in panels A and B. Bar=20 µ m.

    Article Snippet: Membranes were incubated with diluted primary antibodies including rabbit anti-AQP6 polyclonal antibody (pAb, #AQP-006, Alomone Labs, Jerusalem, Israel), rabbit anti-AQP2 pAb (#AQP-002, Alomone labs), rabbit anti-ADR b2 pAb (#Sc-569, Santa Cruz, TX, U.S.A.) and rabbit anti-mAChR m1 pAb (#010, Alomone Labs) for 2 hr at room temperature or at 4°C overnight.

    Techniques: Staining, Modification, Immunohistochemistry