aqp2  (Alomone Labs)


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    Structured Review

    Alomone Labs aqp2
    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, <t>AQP2,</t> AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Aqp2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aqp2/product/Alomone Labs
    Average 90 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    aqp2 - by Bioz Stars, 2022-09
    90/100 stars

    Images

    1) Product Images from "Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus"

    Article Title: Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus

    Journal: Aging and Disease

    doi: 10.14336/AD.2017.1127

    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Figure Legend Snippet: Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P

    Techniques Used: Immunofluorescence, Mouse Assay, Expressing

    2) Product Images from "Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus"

    Article Title: Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus

    Journal: Aging and Disease

    doi: 10.14336/AD.2017.1127

    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Figure Legend Snippet: Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P

    Techniques Used: Immunofluorescence, Mouse Assay, Expressing

    3) Product Images from "Empagliflozin Contributes to Polyuria via Regulation of Sodium Transporters and Water Channels in Diabetic Rat Kidneys"

    Article Title: Empagliflozin Contributes to Polyuria via Regulation of Sodium Transporters and Water Channels in Diabetic Rat Kidneys

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2019.00271

    Empagliflozin increases V2R expression but decreases AQP2 expression in diabetic rat kidneys. (A) Representative immunoblot reacting with anti-V2R, anti-AQP2 and anti-p261-AQP2. (B) Densitometric analysis shows that renal expression of V2R protein in the empagliflozin-treated OLETF group was significantly increased compared with untreated OLETF or lixisenatide-treated OLETF rats. ∗ P
    Figure Legend Snippet: Empagliflozin increases V2R expression but decreases AQP2 expression in diabetic rat kidneys. (A) Representative immunoblot reacting with anti-V2R, anti-AQP2 and anti-p261-AQP2. (B) Densitometric analysis shows that renal expression of V2R protein in the empagliflozin-treated OLETF group was significantly increased compared with untreated OLETF or lixisenatide-treated OLETF rats. ∗ P

    Techniques Used: Expressing

    4) Product Images from "Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus"

    Article Title: Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus

    Journal: Aging and Disease

    doi: 10.14336/AD.2017.1127

    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Figure Legend Snippet: Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P

    Techniques Used: Immunofluorescence, Mouse Assay, Expressing

    5) Product Images from "Intra-renal slow cell-cycle cells contribute to the restoration of kidney tubules injured by ischemia/reperfusion"

    Article Title: Intra-renal slow cell-cycle cells contribute to the restoration of kidney tubules injured by ischemia/reperfusion

    Journal: Anatomy & Cell Biology

    doi: 10.5115/acb.2011.44.3.186

    Numbers and characteristics of BrdU-retaining cells in the kidneys. Twelve hours after birth, mice were administered with 50 mg/kg body weight of BrdU, ip, for 6 days. After a chase period of 8 wk, mouse kidneys were harvested. (A) Kidney sections were immunostained using anti-BrdU antibody, and images were taken at lower magnification. The right column is a high magnification image. (B) BrdU-positive cells were counted in 0.1-mm 2 fields of the superficial cortex, deep cortex, outer medulla, and inner medulla of the kidney (10 fields per kidney). Results are expressed as the mean±SE (n=4). (C) Kidney sections were double-stained with anti-BrdU, -Na/K-ATPase, -Na-K-Cl cotransporter-2 (NKCC2), -aquaporin (AQP)1, or -AQP2 antibody. Images were obtained in the corticomedullary junctions. Arrows indicate BrdU-positive cells. Scale bar=250 µm (A), 50 µm (C).
    Figure Legend Snippet: Numbers and characteristics of BrdU-retaining cells in the kidneys. Twelve hours after birth, mice were administered with 50 mg/kg body weight of BrdU, ip, for 6 days. After a chase period of 8 wk, mouse kidneys were harvested. (A) Kidney sections were immunostained using anti-BrdU antibody, and images were taken at lower magnification. The right column is a high magnification image. (B) BrdU-positive cells were counted in 0.1-mm 2 fields of the superficial cortex, deep cortex, outer medulla, and inner medulla of the kidney (10 fields per kidney). Results are expressed as the mean±SE (n=4). (C) Kidney sections were double-stained with anti-BrdU, -Na/K-ATPase, -Na-K-Cl cotransporter-2 (NKCC2), -aquaporin (AQP)1, or -AQP2 antibody. Images were obtained in the corticomedullary junctions. Arrows indicate BrdU-positive cells. Scale bar=250 µm (A), 50 µm (C).

    Techniques Used: Mouse Assay, Staining

    6) Product Images from "Intra-renal slow cell-cycle cells contribute to the restoration of kidney tubules injured by ischemia/reperfusion"

    Article Title: Intra-renal slow cell-cycle cells contribute to the restoration of kidney tubules injured by ischemia/reperfusion

    Journal: Anatomy & Cell Biology

    doi: 10.5115/acb.2011.44.3.186

    Numbers and characteristics of BrdU-retaining cells in the kidneys. Twelve hours after birth, mice were administered with 50 mg/kg body weight of BrdU, ip, for 6 days. After a chase period of 8 wk, mouse kidneys were harvested. (A) Kidney sections were immunostained using anti-BrdU antibody, and images were taken at lower magnification. The right column is a high magnification image. (B) BrdU-positive cells were counted in 0.1-mm 2 fields of the superficial cortex, deep cortex, outer medulla, and inner medulla of the kidney (10 fields per kidney). Results are expressed as the mean±SE (n=4). (C) Kidney sections were double-stained with anti-BrdU, -Na/K-ATPase, -Na-K-Cl cotransporter-2 (NKCC2), -aquaporin (AQP)1, or -AQP2 antibody. Images were obtained in the corticomedullary junctions. Arrows indicate BrdU-positive cells. Scale bar=250 µm (A), 50 µm (C).
    Figure Legend Snippet: Numbers and characteristics of BrdU-retaining cells in the kidneys. Twelve hours after birth, mice were administered with 50 mg/kg body weight of BrdU, ip, for 6 days. After a chase period of 8 wk, mouse kidneys were harvested. (A) Kidney sections were immunostained using anti-BrdU antibody, and images were taken at lower magnification. The right column is a high magnification image. (B) BrdU-positive cells were counted in 0.1-mm 2 fields of the superficial cortex, deep cortex, outer medulla, and inner medulla of the kidney (10 fields per kidney). Results are expressed as the mean±SE (n=4). (C) Kidney sections were double-stained with anti-BrdU, -Na/K-ATPase, -Na-K-Cl cotransporter-2 (NKCC2), -aquaporin (AQP)1, or -AQP2 antibody. Images were obtained in the corticomedullary junctions. Arrows indicate BrdU-positive cells. Scale bar=250 µm (A), 50 µm (C).

    Techniques Used: Mouse Assay, Staining

    7) Product Images from "Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus"

    Article Title: Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus

    Journal: Aging and Disease

    doi: 10.14336/AD.2017.1127

    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Figure Legend Snippet: Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P

    Techniques Used: Immunofluorescence, Mouse Assay, Expressing

    8) Product Images from "Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus"

    Article Title: Progranulin Deficient Mice Develop Nephrogenic Diabetes Insipidus

    Journal: Aging and Disease

    doi: 10.14336/AD.2017.1127

    Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P
    Figure Legend Snippet: Immunofluorescence analysis of aquaporin 1, 2 and 4 (AQP1, AQP2, AQP4) in the kidney of aged progranulin deficient (Grn -/- ) and control mice (Grn +/+ ) A) Examples of AQP2 immunofluorescence at low (upper panels) and high magnifications (bottom panels and insert). Scale bars as indicated in the figure. The bottom panel also shows high AQP1 and AQP4 at high magnification, which did not differ between genotypes. B) Quantification of AQP2 positive particles using stitched full sections of the kidney of 4 mice per group. Each scatter is a section. Analysis of the tubule lumen as assessed by measuring the lumen area in 4 sections of 4 mice per group. Each scatter is one AQP2 positive collecting duct. The images suggest reduced AQP2 expression and widening of AQP2+ collecting ducts. Asterisks indicate significant differences between genotypes, unpaired 2-tailed Student’s t-test *P

    Techniques Used: Immunofluorescence, Mouse Assay, Expressing

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    Alomone Labs rabbit anti aqp2 antibody
    Mass spectrometry analysis of synthetic peptides corresponding to the COOH-terminal tail of human <t>AQP2</t> did not detect any significant phosphorylation of the water channel by AMPK. Protease-treated peptides (trypsin or LysC) or full-length peptides are indicated with bold underlined amino acids, indicating where phosphorylation was detected. The no. in parentheses indicates the adjusted spectral count of that particular phosphorylation site. The adjusted spectral count is obtained by multiplying the spectral count of each site with its phosphorylated probability reported by PhosphoRS. The last column indicates the percent sequence coverage. SAMS peptide was used as a positive control.
    Rabbit Anti Aqp2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti aqp2 antibody/product/Alomone Labs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rabbit anti aqp2 antibody - by Bioz Stars, 2022-09
    93/100 stars
      Buy from Supplier

    91
    Alomone Labs anti aqp2
    Systemic GSK1016790A treatment normalizes subcellular distribution of TRPV4 and <t>AQP2</t> in freshly isolated CD-derived cyst monolayers. Representative confocal plane micrographs (axes are shown) and corresponding cross-sections (arrows) of three-dimensional
    Anti Aqp2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti aqp2/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti aqp2 - by Bioz Stars, 2022-09
    91/100 stars
      Buy from Supplier

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    Mass spectrometry analysis of synthetic peptides corresponding to the COOH-terminal tail of human AQP2 did not detect any significant phosphorylation of the water channel by AMPK. Protease-treated peptides (trypsin or LysC) or full-length peptides are indicated with bold underlined amino acids, indicating where phosphorylation was detected. The no. in parentheses indicates the adjusted spectral count of that particular phosphorylation site. The adjusted spectral count is obtained by multiplying the spectral count of each site with its phosphorylated probability reported by PhosphoRS. The last column indicates the percent sequence coverage. SAMS peptide was used as a positive control.

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: Mass spectrometry analysis of synthetic peptides corresponding to the COOH-terminal tail of human AQP2 did not detect any significant phosphorylation of the water channel by AMPK. Protease-treated peptides (trypsin or LysC) or full-length peptides are indicated with bold underlined amino acids, indicating where phosphorylation was detected. The no. in parentheses indicates the adjusted spectral count of that particular phosphorylation site. The adjusted spectral count is obtained by multiplying the spectral count of each site with its phosphorylated probability reported by PhosphoRS. The last column indicates the percent sequence coverage. SAMS peptide was used as a positive control.

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Mass Spectrometry, Sequencing, Positive Control

    AMPK activator AICAR prevents apical membrane accumulation of AQP2. Polarized mpkCCD c14 cells were treated with vehicle (DMSO) or the AMPK activator AICAR (1 mM) for 4 h and then 10 μM forskolin (vs. vehicle) for the last 2 h before cell harvesting. 5% of the cell lysates were used for immunoblotting using antibodies against AQP2 or actin as a loading control. The rest of the lysates were affinity purified using streptavidin followed by immunoblotting. A : representative immunoblots of 3 separate surface biotinylation experiments are shown. B : summary densitometric data comparing the mean (±SE) amounts of apical AQP2 as a percentage of that in the control condition for nonglycosylated (solid bars), core glycosylated (open bars), and fully glycosylated (shaded bars) forms of AQP2 are shown. Nonglycosylated: * P

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: AMPK activator AICAR prevents apical membrane accumulation of AQP2. Polarized mpkCCD c14 cells were treated with vehicle (DMSO) or the AMPK activator AICAR (1 mM) for 4 h and then 10 μM forskolin (vs. vehicle) for the last 2 h before cell harvesting. 5% of the cell lysates were used for immunoblotting using antibodies against AQP2 or actin as a loading control. The rest of the lysates were affinity purified using streptavidin followed by immunoblotting. A : representative immunoblots of 3 separate surface biotinylation experiments are shown. B : summary densitometric data comparing the mean (±SE) amounts of apical AQP2 as a percentage of that in the control condition for nonglycosylated (solid bars), core glycosylated (open bars), and fully glycosylated (shaded bars) forms of AQP2 are shown. Nonglycosylated: * P

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Cell Harvesting, Affinity Purification, Western Blot

    AMPK activator AICAR induces intracellular redistribution of AQP2 in ex vivo kidney slices. A : confocal images of AQP2 immunofluorescence labeling (green) in kidney slices incubated in Ringer buffer alone for 75 min display an apical distribution of the water channel. Asterisks indicate the position of the lumen of the collecting ducts. B : addition of 1 mM AICAR for 75 min induced a cytosolic distribution of AQP2. C : incubation of kidney slices with dDAVP for the last 15 min of a 75-min incubation revealed apical accumulation of AQP2. D : however, preincubation of slices with AICAR did not prevent the dDAVP-induced apical accumulation of AQP2. E : regions of interest (ROIs; example shown in A ). Quantification of the mean (±SE) AQP2-associated mean pixel intensity (MPI) of apical-to-cytoplasmic ratio [arbitrary units (AU)] relative to that of cells measured under the control condition was used as a measure of apical AQP2 accumulation. Additional incubations of kidney slices in Ringer buffer were performed in the absence ( F ) or presence ( G ) of the PKA inhibitor myristoylated protein kinase inhibitor (mPKI; 10 μM) for 75 min. H : the relative AQP2 mean pixel intensity apical-to-cytoplasmic ratio was reduced dramatically with mPKI treatment. Data were obtained from at least 3 separate kidney slice experiments, using kidneys from at least 3 animals, and measuring a total of at least 30 cells per condition. * P

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: AMPK activator AICAR induces intracellular redistribution of AQP2 in ex vivo kidney slices. A : confocal images of AQP2 immunofluorescence labeling (green) in kidney slices incubated in Ringer buffer alone for 75 min display an apical distribution of the water channel. Asterisks indicate the position of the lumen of the collecting ducts. B : addition of 1 mM AICAR for 75 min induced a cytosolic distribution of AQP2. C : incubation of kidney slices with dDAVP for the last 15 min of a 75-min incubation revealed apical accumulation of AQP2. D : however, preincubation of slices with AICAR did not prevent the dDAVP-induced apical accumulation of AQP2. E : regions of interest (ROIs; example shown in A ). Quantification of the mean (±SE) AQP2-associated mean pixel intensity (MPI) of apical-to-cytoplasmic ratio [arbitrary units (AU)] relative to that of cells measured under the control condition was used as a measure of apical AQP2 accumulation. Additional incubations of kidney slices in Ringer buffer were performed in the absence ( F ) or presence ( G ) of the PKA inhibitor myristoylated protein kinase inhibitor (mPKI; 10 μM) for 75 min. H : the relative AQP2 mean pixel intensity apical-to-cytoplasmic ratio was reduced dramatically with mPKI treatment. Data were obtained from at least 3 separate kidney slice experiments, using kidneys from at least 3 animals, and measuring a total of at least 30 cells per condition. * P

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Ex Vivo, Immunofluorescence, Labeling, Incubation

    AMPK inhibition promotes AQP2-mediated oocyte swelling and shortens time to lysis. Oocytes were microinjected with cRNAs for AQP2 and the different indicated AMPK constructs [WT-AMPK-α (●), dominant-negative (DN) AMPK-α1-K45R (■), and constitutively active (CA) AMPK-γ1-R70Q (▲)]. After 3 days, the oocytes were subjected to hypotonic shock in deionized water. Oocyte times to lysis were assessed by microscopy, and the mean times to lysis (normalized to that of the WT-AMPK-expressing group for each batch) are shown for all of the data obtained for these 3 groups. Oocytes expressing DN-AMPK had a significantly reduced time to lysis relative to each of the other two groups. * P

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: AMPK inhibition promotes AQP2-mediated oocyte swelling and shortens time to lysis. Oocytes were microinjected with cRNAs for AQP2 and the different indicated AMPK constructs [WT-AMPK-α (●), dominant-negative (DN) AMPK-α1-K45R (■), and constitutively active (CA) AMPK-γ1-R70Q (▲)]. After 3 days, the oocytes were subjected to hypotonic shock in deionized water. Oocyte times to lysis were assessed by microscopy, and the mean times to lysis (normalized to that of the WT-AMPK-expressing group for each batch) are shown for all of the data obtained for these 3 groups. Oocytes expressing DN-AMPK had a significantly reduced time to lysis relative to each of the other two groups. * P

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Inhibition, Lysis, Construct, Dominant Negative Mutation, Microscopy, Expressing

    AMPK fails to significantly phosphorylate AQP2 in vitro. V5-tagged AQP2 was expressed in HEK-293 cells, immunoprecipitated, and incubated with [γ- 32 P]ATP in the presence of PKA catalytic subunit (positive control) or in the presence or absence of AMPK holoenzyme. A phosphoscreen image ( top ) and immunoblot ( bottom ) of the same membrane are shown (representative of 4 experiments). AQP2 gets robustly phosphorylated in the presence of PKA, but only weakly phosphorylated in the presence of AMPK ( top ). PKA catalytic subunit and the AMPK β-subunit get autophosphorylated, as indicated. The immunoblot ( bottom ) reveals similar AQP2 loading in all lanes.

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: AMPK fails to significantly phosphorylate AQP2 in vitro. V5-tagged AQP2 was expressed in HEK-293 cells, immunoprecipitated, and incubated with [γ- 32 P]ATP in the presence of PKA catalytic subunit (positive control) or in the presence or absence of AMPK holoenzyme. A phosphoscreen image ( top ) and immunoblot ( bottom ) of the same membrane are shown (representative of 4 experiments). AQP2 gets robustly phosphorylated in the presence of PKA, but only weakly phosphorylated in the presence of AMPK ( top ). PKA catalytic subunit and the AMPK β-subunit get autophosphorylated, as indicated. The immunoblot ( bottom ) reveals similar AQP2 loading in all lanes.

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: In Vitro, Immunoprecipitation, Incubation, Positive Control

    Changes in AQP2 phosphorylation pattern following dDAVP treatment in the presence or absence of AMPK activator in mpkCCD c14 cells. Twenty-four hours posttransfection with wild-type AQP2 plasmid, cells were incubated in the presence or absence of AICAR (2 mM) for 4 h, and then dDAVP (10 nM) or vehicle was added for 30 min before cell harvesting. Immunoblot analysis of immunoprecipitated V5-AQP2 was performed using the corresponding rabbit polyclonal antibodies against phosphorylated AQP2 COOH-terminal sites: −S256, −S261, −S264, and −S269. To confirm successful immunoprecipitation, the membranes were reblotted using anti-AQP2 and anti-V5 antibodies coupled to HRP. A : representative set of immunoblots from these experiments using antibodies against AQP2 and its different COOH-terminal phosphorylation sites. B : quantification of AQP2 phosphorylation signal for different AQP2 COOH-terminal sites normalized for protein loading, as assessed by densitometry of the immunoblot. Values are means ± SE of 4 independent experiments. * P

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of the metabolic sensor AMP-activated protein kinase inhibits aquaporin-2 function in kidney principal cells

    doi: 10.1152/ajprenal.00308.2016

    Figure Lengend Snippet: Changes in AQP2 phosphorylation pattern following dDAVP treatment in the presence or absence of AMPK activator in mpkCCD c14 cells. Twenty-four hours posttransfection with wild-type AQP2 plasmid, cells were incubated in the presence or absence of AICAR (2 mM) for 4 h, and then dDAVP (10 nM) or vehicle was added for 30 min before cell harvesting. Immunoblot analysis of immunoprecipitated V5-AQP2 was performed using the corresponding rabbit polyclonal antibodies against phosphorylated AQP2 COOH-terminal sites: −S256, −S261, −S264, and −S269. To confirm successful immunoprecipitation, the membranes were reblotted using anti-AQP2 and anti-V5 antibodies coupled to HRP. A : representative set of immunoblots from these experiments using antibodies against AQP2 and its different COOH-terminal phosphorylation sites. B : quantification of AQP2 phosphorylation signal for different AQP2 COOH-terminal sites normalized for protein loading, as assessed by densitometry of the immunoblot. Values are means ± SE of 4 independent experiments. * P

    Article Snippet: The rabbit anti-AQP2 antibody was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Plasmid Preparation, Incubation, Cell Harvesting, Immunoprecipitation, Western Blot

    Downregulation of AQP2 mRNA by LiCl is time and concentration dependent. IMCD cells were treated for different time points with LiCl (20 mM). The mRNA expression of AQP2-4, BGT1, and AR was analyzed by real-time PCR and the relative changes compared to untreated cells were calculated ( a ). In the same way, IMCD cells were treated for 24 h with 10 or 20 mM of LiCl and the relative changes in the gene expression of AQP2 and AR were compared to untreated cells ( b ). One-way ANOVA analysis with Tukey post-test, * indicates statistically significant differences to untreated cells ( p ≤ 0.05); n = 3.

    Journal: Cells

    Article Title: Lithium Chloride and GSK3 Inhibition Reduce Aquaporin-2 Expression in Primary Cultured Inner Medullary Collecting Duct Cells Due to Independent Mechanisms

    doi: 10.3390/cells9041060

    Figure Lengend Snippet: Downregulation of AQP2 mRNA by LiCl is time and concentration dependent. IMCD cells were treated for different time points with LiCl (20 mM). The mRNA expression of AQP2-4, BGT1, and AR was analyzed by real-time PCR and the relative changes compared to untreated cells were calculated ( a ). In the same way, IMCD cells were treated for 24 h with 10 or 20 mM of LiCl and the relative changes in the gene expression of AQP2 and AR were compared to untreated cells ( b ). One-way ANOVA analysis with Tukey post-test, * indicates statistically significant differences to untreated cells ( p ≤ 0.05); n = 3.

    Article Snippet: Antibodies raised against Aqp2-4 were obtained from Alomone Labs (Jerusalem, Israel) and antibodies directed against phosphorylated Aqp2 at position S256 were a generous gift from M. Knepper (National Institutes of Health, Bethesda, MD, USA).

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

    MG132 and bafilomycin induce downregulation of Aqp2 on the mRNA level. IMCD cells were treated for 1 or 24 h with bafilomycin (Baf. 100 nM) or MG132 (2 µM) and the mRNA expression of Aqp2-4, BGT-1, and AR was compared to untreated cells by real-time PCR. One-way ANOVA with tukey post-test was performed and * indicates statistically significant differences compared untreated cells ( p ≤ 0.05).

    Journal: Cells

    Article Title: Lithium Chloride and GSK3 Inhibition Reduce Aquaporin-2 Expression in Primary Cultured Inner Medullary Collecting Duct Cells Due to Independent Mechanisms

    doi: 10.3390/cells9041060

    Figure Lengend Snippet: MG132 and bafilomycin induce downregulation of Aqp2 on the mRNA level. IMCD cells were treated for 1 or 24 h with bafilomycin (Baf. 100 nM) or MG132 (2 µM) and the mRNA expression of Aqp2-4, BGT-1, and AR was compared to untreated cells by real-time PCR. One-way ANOVA with tukey post-test was performed and * indicates statistically significant differences compared untreated cells ( p ≤ 0.05).

    Article Snippet: Antibodies raised against Aqp2-4 were obtained from Alomone Labs (Jerusalem, Israel) and antibodies directed against phosphorylated Aqp2 at position S256 were a generous gift from M. Knepper (National Institutes of Health, Bethesda, MD, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Migration influences the localization of AQPs. Wound healing assays were performed with IMCD-cells cultivated at 600 mosmol/kg. A scratch was induced and after 4 h the cells were stained for AQP2–4 using specific antibodies. For the AQP2 staining the cells were incubated with dbcAMP to induce membrane localization. Alexa-488 labeled secondary antibodies were used for visualization. The nuclei were stained with DAPI. The upper row show cells away from the wound. The lower rows show cells at the leading edge. Scale bar = 20 µm, representative image chosen from 6 to 9 independent experiments.

    Journal: Scientific Reports

    Article Title: Unexpected localization of AQP3 and AQP4 induced by migration of primary cultured IMCD cells

    doi: 10.1038/s41598-021-91369-y

    Figure Lengend Snippet: Migration influences the localization of AQPs. Wound healing assays were performed with IMCD-cells cultivated at 600 mosmol/kg. A scratch was induced and after 4 h the cells were stained for AQP2–4 using specific antibodies. For the AQP2 staining the cells were incubated with dbcAMP to induce membrane localization. Alexa-488 labeled secondary antibodies were used for visualization. The nuclei were stained with DAPI. The upper row show cells away from the wound. The lower rows show cells at the leading edge. Scale bar = 20 µm, representative image chosen from 6 to 9 independent experiments.

    Article Snippet: Antibodies raised against AQP2–4 were obtained from Alomone Labs (Jerusalem, Israel) .

    Techniques: Migration, Staining, Incubation, Labeling

    Systemic GSK1016790A treatment normalizes subcellular distribution of TRPV4 and AQP2 in freshly isolated CD-derived cyst monolayers. Representative confocal plane micrographs (axes are shown) and corresponding cross-sections (arrows) of three-dimensional

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: TRPV4 Dysfunction Promotes Renal Cystogenesis in Autosomal Recessive Polycystic Kidney Disease

    doi: 10.1681/ASN.2012050442

    Figure Lengend Snippet: Systemic GSK1016790A treatment normalizes subcellular distribution of TRPV4 and AQP2 in freshly isolated CD-derived cyst monolayers. Representative confocal plane micrographs (axes are shown) and corresponding cross-sections (arrows) of three-dimensional

    Article Snippet: After washing with PBS (three times for 5 minutes) the samples were incubated for 3 hours at RT in dark with the mix of anti-TRPV4 (1:500; Alomone Labs), anti-AQP2 tagged with ATTO 550 (1:100; Alomone Labs) in 1% serum + 0.1% Triton in PBS.

    Techniques: Isolation, Derivative Assay