rabbit anti aqp1  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti aqp1
    <t>AQP1</t> and NKCC1 are present in leptomeningeal vascular endothelia of the spinal cord. Micrographs of paraffin sections obtained from the spinal cord of adult mice (P90) and immunolabeled for AQP1 and NKCC1 (brown). AQP1 immunoreactivity is predominantly located in C fibers in the dorsal horns of the spinal cord ( a , arrowheads), whereas NKCC1 is observed throughout the spinal cord grey matter ( d ). b , e High magnification of the area delimited by the blue rectangle in a and d , respectively, show AQP1int + /NKCC1 + leptomeningeal vessels (arrows) in the spinal cord. c , f High magnification micrographs of the area delimited by the green squares in b and e show AQP1int + /NKCC1 + cells in the vascular endothelium, restricted to the subarachnoid space along the spinal cord (curved arrowheads). DRG, dorsal root ganglia; SAS, subarachnoid space. Scale bars: a , d 1 mm; b , e 100 μm; c , f 50 μm
    Rabbit Anti Aqp1, 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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    rabbit anti aqp1 - by Bioz Stars, 2022-01
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    Images

    1) Product Images from "Aquaporin 1 and the Na+/K+/2Cl− cotransporter 1 are present in the leptomeningeal vasculature of the adult rodent central nervous system"

    Article Title: Aquaporin 1 and the Na+/K+/2Cl− cotransporter 1 are present in the leptomeningeal vasculature of the adult rodent central nervous system

    Journal: Fluids and Barriers of the CNS

    doi: 10.1186/s12987-020-0176-z

    AQP1 and NKCC1 are present in leptomeningeal vascular endothelia of the spinal cord. Micrographs of paraffin sections obtained from the spinal cord of adult mice (P90) and immunolabeled for AQP1 and NKCC1 (brown). AQP1 immunoreactivity is predominantly located in C fibers in the dorsal horns of the spinal cord ( a , arrowheads), whereas NKCC1 is observed throughout the spinal cord grey matter ( d ). b , e High magnification of the area delimited by the blue rectangle in a and d , respectively, show AQP1int + /NKCC1 + leptomeningeal vessels (arrows) in the spinal cord. c , f High magnification micrographs of the area delimited by the green squares in b and e show AQP1int + /NKCC1 + cells in the vascular endothelium, restricted to the subarachnoid space along the spinal cord (curved arrowheads). DRG, dorsal root ganglia; SAS, subarachnoid space. Scale bars: a , d 1 mm; b , e 100 μm; c , f 50 μm
    Figure Legend Snippet: AQP1 and NKCC1 are present in leptomeningeal vascular endothelia of the spinal cord. Micrographs of paraffin sections obtained from the spinal cord of adult mice (P90) and immunolabeled for AQP1 and NKCC1 (brown). AQP1 immunoreactivity is predominantly located in C fibers in the dorsal horns of the spinal cord ( a , arrowheads), whereas NKCC1 is observed throughout the spinal cord grey matter ( d ). b , e High magnification of the area delimited by the blue rectangle in a and d , respectively, show AQP1int + /NKCC1 + leptomeningeal vessels (arrows) in the spinal cord. c , f High magnification micrographs of the area delimited by the green squares in b and e show AQP1int + /NKCC1 + cells in the vascular endothelium, restricted to the subarachnoid space along the spinal cord (curved arrowheads). DRG, dorsal root ganglia; SAS, subarachnoid space. Scale bars: a , d 1 mm; b , e 100 μm; c , f 50 μm

    Techniques Used: Mouse Assay, Immunolabeling

    AQP1 and NKCC1 distribution in the CNS leptomeningeal vasculature. Scheme representing the mouse brain parenchyma, the skull and the meninges, which encompass the brain and also the spinal cord. The meninges are divided into the dura mater and the leptomeninges, corresponding to the arachnoid and pia mater. The brain and spinal parenchyma are separated from the meninges by the basal lamina and the glia limitans. The arachnoid mater forms the outer barrier of the CNS and underneath it lies the subarachnoid space (SAS), which is filled with CSF. Immune cells, namely macrophages and leucocytes, are sparsely present within the SAS, surveilling the healthy CNS. Additionally to its function as route for CSF and immune cells circulation, the SAS encloses the arterial blood supply to the CNS. Prior to entering the CNS parenchyma, leptomeningeal arteries branch and divide into arterioles. Within the parenchyma, penetrating arterioles and veins are tethered by astrocytes with highly polarized AQP4 distribution, a unique feature of the CNS vasculature. Schematic representation of cross sections of the leptomeningeal vasculature denotes AQP1 and NKCC1 expression by smooth muscle cells, which compose the tunica media of arterioles and veins. In contrast, endothelial cells within the tunica intima are devoid of both proteins. Notwithstanding, endothelial cells of capillaries and venules present both AQP1 and NKCC1
    Figure Legend Snippet: AQP1 and NKCC1 distribution in the CNS leptomeningeal vasculature. Scheme representing the mouse brain parenchyma, the skull and the meninges, which encompass the brain and also the spinal cord. The meninges are divided into the dura mater and the leptomeninges, corresponding to the arachnoid and pia mater. The brain and spinal parenchyma are separated from the meninges by the basal lamina and the glia limitans. The arachnoid mater forms the outer barrier of the CNS and underneath it lies the subarachnoid space (SAS), which is filled with CSF. Immune cells, namely macrophages and leucocytes, are sparsely present within the SAS, surveilling the healthy CNS. Additionally to its function as route for CSF and immune cells circulation, the SAS encloses the arterial blood supply to the CNS. Prior to entering the CNS parenchyma, leptomeningeal arteries branch and divide into arterioles. Within the parenchyma, penetrating arterioles and veins are tethered by astrocytes with highly polarized AQP4 distribution, a unique feature of the CNS vasculature. Schematic representation of cross sections of the leptomeningeal vasculature denotes AQP1 and NKCC1 expression by smooth muscle cells, which compose the tunica media of arterioles and veins. In contrast, endothelial cells within the tunica intima are devoid of both proteins. Notwithstanding, endothelial cells of capillaries and venules present both AQP1 and NKCC1

    Techniques Used: Expressing

    uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm
    Figure Legend Snippet: uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm

    Techniques Used: Expressing, Immunolabeling, Mouse Assay, Binding Assay

    AQP1 is expressed in the brain and peripheral vasculature. a Confocal micrograph from an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, magenta and AQP1int, green). DAPI nuclear counterstaining (blue). b AQP1ext + blood vessel, located around the ventricles (delimited by the magenta square in a ). c – f Immunoreactive epithelial choroid plexus cells, located in the lateral ventricles, are labeled with both antibodies (high magnification of the area delimited by the green square in a ). g , h Micrographs of mouse kidney show the distribution of AQP1 in the vascular endothelium and proximal tubules. i , j Higher magnification image of a blood vessel immunolabeled for CD31 (green) and AQP1int (magenta) (delimited by square in h ). Asterisk indicates the lumen of a blood vessel and arrows indicate proximal tubules. k , l AQP1 + endothelial cells are also detected in the heart of adult mice. m – o Paraffin sections obtained from adult rat brain show AQP1 immunoreactive blood vessels in the hippocampal fissure and epithelial cells of the choroid plexus located in the third ventricle. Arrows and curved arrowheads indicate arterioles or veins and capillaries or venules, respectively. Straight arrowheads indicate AQP1 − blood vessels. 3V, third ventricle; BV, blood vessel; ChP, choroid plexus; DG, dentate gyrus; LV, lateral ventricle; PT, proximal tubule. Scale bars: a , b and g – j 50 µm; c – f 5 µm; k 1 mm; l 100 µm; m 2 mm; n 500 μm; o 200 μm
    Figure Legend Snippet: AQP1 is expressed in the brain and peripheral vasculature. a Confocal micrograph from an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, magenta and AQP1int, green). DAPI nuclear counterstaining (blue). b AQP1ext + blood vessel, located around the ventricles (delimited by the magenta square in a ). c – f Immunoreactive epithelial choroid plexus cells, located in the lateral ventricles, are labeled with both antibodies (high magnification of the area delimited by the green square in a ). g , h Micrographs of mouse kidney show the distribution of AQP1 in the vascular endothelium and proximal tubules. i , j Higher magnification image of a blood vessel immunolabeled for CD31 (green) and AQP1int (magenta) (delimited by square in h ). Asterisk indicates the lumen of a blood vessel and arrows indicate proximal tubules. k , l AQP1 + endothelial cells are also detected in the heart of adult mice. m – o Paraffin sections obtained from adult rat brain show AQP1 immunoreactive blood vessels in the hippocampal fissure and epithelial cells of the choroid plexus located in the third ventricle. Arrows and curved arrowheads indicate arterioles or veins and capillaries or venules, respectively. Straight arrowheads indicate AQP1 − blood vessels. 3V, third ventricle; BV, blood vessel; ChP, choroid plexus; DG, dentate gyrus; LV, lateral ventricle; PT, proximal tubule. Scale bars: a , b and g – j 50 µm; c – f 5 µm; k 1 mm; l 100 µm; m 2 mm; n 500 μm; o 200 μm

    Techniques Used: Immunolabeling, Labeling, Mouse Assay

    AQP1 and NKCC1 are present in smooth muscle and endothelial cells of the leptomeningeal vasculature. a , b Paraffin sections of adult mouse brain (P90) immunolabeled with anti-AQP1int or anti-NKCC1 (both brown). c Some sections were stained with hematoxylin (HE, pink) and the vascular identity of blood vessels located in the subarachnoid space (cisterna interpendicularis, delimited by square in a , b ) was determined. d , e Consecutive sections show that AQP1int + /NKCC1 + cells are present in the smooth muscle cell layer of arterioles (arrowheads) and in the endothelium of capillaries and venules, respectively (curved arrowheads). f , g Vascular endothelial cells were labeled by lectin (WGA-FITC, green), followed by standard Immunolabeling. DAPI counterstain (blue) reveal the location of the leptomeningeal vessel (asterisk). h – j Higher magnification confocal images show that AQP1 is restricted to tunica media, where AQP1ext + smooth muscle cells, identified by their round soma (arrowheads) are observed, whereas AQP1 is not present in the endothelial cell layer (curved arrowheads). The arrow indicates a leptomeningeal cell, also AQP1ext + . BS, brain stem; Cb, cerebellum; cp, cerebral peduncle; Ctx, cerebral cortex; Hip, hippocampus; Hyp, hypothalamus; OB, olfactory bulb; Pn, pontine nuclei. Scale bars: a , b 2 mm; c – e 100 μm; f – j 50 μm
    Figure Legend Snippet: AQP1 and NKCC1 are present in smooth muscle and endothelial cells of the leptomeningeal vasculature. a , b Paraffin sections of adult mouse brain (P90) immunolabeled with anti-AQP1int or anti-NKCC1 (both brown). c Some sections were stained with hematoxylin (HE, pink) and the vascular identity of blood vessels located in the subarachnoid space (cisterna interpendicularis, delimited by square in a , b ) was determined. d , e Consecutive sections show that AQP1int + /NKCC1 + cells are present in the smooth muscle cell layer of arterioles (arrowheads) and in the endothelium of capillaries and venules, respectively (curved arrowheads). f , g Vascular endothelial cells were labeled by lectin (WGA-FITC, green), followed by standard Immunolabeling. DAPI counterstain (blue) reveal the location of the leptomeningeal vessel (asterisk). h – j Higher magnification confocal images show that AQP1 is restricted to tunica media, where AQP1ext + smooth muscle cells, identified by their round soma (arrowheads) are observed, whereas AQP1 is not present in the endothelial cell layer (curved arrowheads). The arrow indicates a leptomeningeal cell, also AQP1ext + . BS, brain stem; Cb, cerebellum; cp, cerebral peduncle; Ctx, cerebral cortex; Hip, hippocampus; Hyp, hypothalamus; OB, olfactory bulb; Pn, pontine nuclei. Scale bars: a , b 2 mm; c – e 100 μm; f – j 50 μm

    Techniques Used: Immunolabeling, Staining, Labeling, Whole Genome Amplification

    AQP1 and NKCC1 are expressed by the choroidal epithelial cells and in the leptomeningeal vasculature. a – f Confocal micrograph show a leptomeningeal WGA-FITC + (green) labeled vessel immunoreactive for AQP1 (magenta) and NKCC1 (orange) in the adult mouse brain (P90). In b an optical section reveals that AQP1 + /NKCC1 + cells are restricted to the smooth muscle cell layer (arrowheads) and absent in the endothelial cells (curved arrowheads), which are labeled by WGA-FITC. g , h NKCC1 is detected in the choroid plexus epithelia, in ependymal cells and in the molecular layer of the cerebellum, as shown in the micrographs of the fourth ventricle. i Double labeling confirms AQP1 and NKCC1 presence in choroid plexus epithelial cells (higher magnification of the area delimited by the blue square in h ). j , k Brain sections obtained from NKCC1 KO adult mice show no immunoreactivity in the brain parenchyma neither in the choroid plexus. l , m Histological sections immunolabeled with antibodies against AQP1ext (magenta), NKCC1 (yellow) and α-SMA (cyan), reveal AQP1ext + /NKCC1 + /α-SMA + leptomeningeal vessels around the hippocampus and third ventricle. Low magnification micrograph shows DAPI (blue) counterstaining and indicates a leptomeningeal blood vessel (asterisk) closely located to the hippocampal fissure. n – p Higher magnification of an AQP1ext + /NKCC1 + vessel (delimited by the dashed square in j . Arrowheads indicate α-SMA + cells. ( q ) Optical sectioning reveals that both AQP1 and NKCC1 are distributed in the smooth muscle cell layer (arrowheads). r 3D rendering of the leptomeningeal vessel confirms AQP1 and NKCC1 restriction to the smooth muscle cell layer (arrowheads). ChP, choroid plexus; DG, dentate gyrus; DS, dorsal subiculum; GL, granular layer; hif, hippocampal fissure; Mol, molecular layer; SAS, subarachnoid space; 3V, third ventricle, 4V, fourth ventricle. Scale bars: a , i 20 µm; b – f , q , r 10 µm; g , h , j – p 50 µm
    Figure Legend Snippet: AQP1 and NKCC1 are expressed by the choroidal epithelial cells and in the leptomeningeal vasculature. a – f Confocal micrograph show a leptomeningeal WGA-FITC + (green) labeled vessel immunoreactive for AQP1 (magenta) and NKCC1 (orange) in the adult mouse brain (P90). In b an optical section reveals that AQP1 + /NKCC1 + cells are restricted to the smooth muscle cell layer (arrowheads) and absent in the endothelial cells (curved arrowheads), which are labeled by WGA-FITC. g , h NKCC1 is detected in the choroid plexus epithelia, in ependymal cells and in the molecular layer of the cerebellum, as shown in the micrographs of the fourth ventricle. i Double labeling confirms AQP1 and NKCC1 presence in choroid plexus epithelial cells (higher magnification of the area delimited by the blue square in h ). j , k Brain sections obtained from NKCC1 KO adult mice show no immunoreactivity in the brain parenchyma neither in the choroid plexus. l , m Histological sections immunolabeled with antibodies against AQP1ext (magenta), NKCC1 (yellow) and α-SMA (cyan), reveal AQP1ext + /NKCC1 + /α-SMA + leptomeningeal vessels around the hippocampus and third ventricle. Low magnification micrograph shows DAPI (blue) counterstaining and indicates a leptomeningeal blood vessel (asterisk) closely located to the hippocampal fissure. n – p Higher magnification of an AQP1ext + /NKCC1 + vessel (delimited by the dashed square in j . Arrowheads indicate α-SMA + cells. ( q ) Optical sectioning reveals that both AQP1 and NKCC1 are distributed in the smooth muscle cell layer (arrowheads). r 3D rendering of the leptomeningeal vessel confirms AQP1 and NKCC1 restriction to the smooth muscle cell layer (arrowheads). ChP, choroid plexus; DG, dentate gyrus; DS, dorsal subiculum; GL, granular layer; hif, hippocampal fissure; Mol, molecular layer; SAS, subarachnoid space; 3V, third ventricle, 4V, fourth ventricle. Scale bars: a , i 20 µm; b – f , q , r 10 µm; g , h , j – p 50 µm

    Techniques Used: Whole Genome Amplification, Labeling, Mouse Assay, Immunolabeling

    2) Product Images from "Aquaporin 1 and the Na+/K+/2Cl− cotransporter 1 are present in the leptomeningeal vasculature of the adult rodent central nervous system"

    Article Title: Aquaporin 1 and the Na+/K+/2Cl− cotransporter 1 are present in the leptomeningeal vasculature of the adult rodent central nervous system

    Journal: Fluids and Barriers of the CNS

    doi: 10.1186/s12987-020-0176-z

    AQP1 and NKCC1 are present in leptomeningeal vascular endothelia of the spinal cord. Micrographs of paraffin sections obtained from the spinal cord of adult mice (P90) and immunolabeled for AQP1 and NKCC1 (brown). AQP1 immunoreactivity is predominantly located in C fibers in the dorsal horns of the spinal cord ( a , arrowheads), whereas NKCC1 is observed throughout the spinal cord grey matter ( d ). b , e High magnification of the area delimited by the blue rectangle in a and d , respectively, show AQP1int + /NKCC1 + leptomeningeal vessels (arrows) in the spinal cord. c , f High magnification micrographs of the area delimited by the green squares in b and e show AQP1int + /NKCC1 + cells in the vascular endothelium, restricted to the subarachnoid space along the spinal cord (curved arrowheads). DRG, dorsal root ganglia; SAS, subarachnoid space. Scale bars: a , d 1 mm; b , e 100 μm; c , f 50 μm
    Figure Legend Snippet: AQP1 and NKCC1 are present in leptomeningeal vascular endothelia of the spinal cord. Micrographs of paraffin sections obtained from the spinal cord of adult mice (P90) and immunolabeled for AQP1 and NKCC1 (brown). AQP1 immunoreactivity is predominantly located in C fibers in the dorsal horns of the spinal cord ( a , arrowheads), whereas NKCC1 is observed throughout the spinal cord grey matter ( d ). b , e High magnification of the area delimited by the blue rectangle in a and d , respectively, show AQP1int + /NKCC1 + leptomeningeal vessels (arrows) in the spinal cord. c , f High magnification micrographs of the area delimited by the green squares in b and e show AQP1int + /NKCC1 + cells in the vascular endothelium, restricted to the subarachnoid space along the spinal cord (curved arrowheads). DRG, dorsal root ganglia; SAS, subarachnoid space. Scale bars: a , d 1 mm; b , e 100 μm; c , f 50 μm

    Techniques Used: Mouse Assay, Immunolabeling

    AQP1 and NKCC1 distribution in the CNS leptomeningeal vasculature. Scheme representing the mouse brain parenchyma, the skull and the meninges, which encompass the brain and also the spinal cord. The meninges are divided into the dura mater and the leptomeninges, corresponding to the arachnoid and pia mater. The brain and spinal parenchyma are separated from the meninges by the basal lamina and the glia limitans. The arachnoid mater forms the outer barrier of the CNS and underneath it lies the subarachnoid space (SAS), which is filled with CSF. Immune cells, namely macrophages and leucocytes, are sparsely present within the SAS, surveilling the healthy CNS. Additionally to its function as route for CSF and immune cells circulation, the SAS encloses the arterial blood supply to the CNS. Prior to entering the CNS parenchyma, leptomeningeal arteries branch and divide into arterioles. Within the parenchyma, penetrating arterioles and veins are tethered by astrocytes with highly polarized AQP4 distribution, a unique feature of the CNS vasculature. Schematic representation of cross sections of the leptomeningeal vasculature denotes AQP1 and NKCC1 expression by smooth muscle cells, which compose the tunica media of arterioles and veins. In contrast, endothelial cells within the tunica intima are devoid of both proteins. Notwithstanding, endothelial cells of capillaries and venules present both AQP1 and NKCC1
    Figure Legend Snippet: AQP1 and NKCC1 distribution in the CNS leptomeningeal vasculature. Scheme representing the mouse brain parenchyma, the skull and the meninges, which encompass the brain and also the spinal cord. The meninges are divided into the dura mater and the leptomeninges, corresponding to the arachnoid and pia mater. The brain and spinal parenchyma are separated from the meninges by the basal lamina and the glia limitans. The arachnoid mater forms the outer barrier of the CNS and underneath it lies the subarachnoid space (SAS), which is filled with CSF. Immune cells, namely macrophages and leucocytes, are sparsely present within the SAS, surveilling the healthy CNS. Additionally to its function as route for CSF and immune cells circulation, the SAS encloses the arterial blood supply to the CNS. Prior to entering the CNS parenchyma, leptomeningeal arteries branch and divide into arterioles. Within the parenchyma, penetrating arterioles and veins are tethered by astrocytes with highly polarized AQP4 distribution, a unique feature of the CNS vasculature. Schematic representation of cross sections of the leptomeningeal vasculature denotes AQP1 and NKCC1 expression by smooth muscle cells, which compose the tunica media of arterioles and veins. In contrast, endothelial cells within the tunica intima are devoid of both proteins. Notwithstanding, endothelial cells of capillaries and venules present both AQP1 and NKCC1

    Techniques Used: Expressing

    uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm
    Figure Legend Snippet: uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm

    Techniques Used: Expressing, Immunolabeling, Mouse Assay, Binding Assay

    AQP1 is expressed in the brain and peripheral vasculature. a Confocal micrograph from an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, magenta and AQP1int, green). DAPI nuclear counterstaining (blue). b AQP1ext + blood vessel, located around the ventricles (delimited by the magenta square in a ). c – f Immunoreactive epithelial choroid plexus cells, located in the lateral ventricles, are labeled with both antibodies (high magnification of the area delimited by the green square in a ). g , h Micrographs of mouse kidney show the distribution of AQP1 in the vascular endothelium and proximal tubules. i , j Higher magnification image of a blood vessel immunolabeled for CD31 (green) and AQP1int (magenta) (delimited by square in h ). Asterisk indicates the lumen of a blood vessel and arrows indicate proximal tubules. k , l AQP1 + endothelial cells are also detected in the heart of adult mice. m – o Paraffin sections obtained from adult rat brain show AQP1 immunoreactive blood vessels in the hippocampal fissure and epithelial cells of the choroid plexus located in the third ventricle. Arrows and curved arrowheads indicate arterioles or veins and capillaries or venules, respectively. Straight arrowheads indicate AQP1 − blood vessels. 3V, third ventricle; BV, blood vessel; ChP, choroid plexus; DG, dentate gyrus; LV, lateral ventricle; PT, proximal tubule. Scale bars: a , b and g – j 50 µm; c – f 5 µm; k 1 mm; l 100 µm; m 2 mm; n 500 μm; o 200 μm
    Figure Legend Snippet: AQP1 is expressed in the brain and peripheral vasculature. a Confocal micrograph from an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, magenta and AQP1int, green). DAPI nuclear counterstaining (blue). b AQP1ext + blood vessel, located around the ventricles (delimited by the magenta square in a ). c – f Immunoreactive epithelial choroid plexus cells, located in the lateral ventricles, are labeled with both antibodies (high magnification of the area delimited by the green square in a ). g , h Micrographs of mouse kidney show the distribution of AQP1 in the vascular endothelium and proximal tubules. i , j Higher magnification image of a blood vessel immunolabeled for CD31 (green) and AQP1int (magenta) (delimited by square in h ). Asterisk indicates the lumen of a blood vessel and arrows indicate proximal tubules. k , l AQP1 + endothelial cells are also detected in the heart of adult mice. m – o Paraffin sections obtained from adult rat brain show AQP1 immunoreactive blood vessels in the hippocampal fissure and epithelial cells of the choroid plexus located in the third ventricle. Arrows and curved arrowheads indicate arterioles or veins and capillaries or venules, respectively. Straight arrowheads indicate AQP1 − blood vessels. 3V, third ventricle; BV, blood vessel; ChP, choroid plexus; DG, dentate gyrus; LV, lateral ventricle; PT, proximal tubule. Scale bars: a , b and g – j 50 µm; c – f 5 µm; k 1 mm; l 100 µm; m 2 mm; n 500 μm; o 200 μm

    Techniques Used: Immunolabeling, Labeling, Mouse Assay

    AQP1 and NKCC1 are present in smooth muscle and endothelial cells of the leptomeningeal vasculature. a , b Paraffin sections of adult mouse brain (P90) immunolabeled with anti-AQP1int or anti-NKCC1 (both brown). c Some sections were stained with hematoxylin (HE, pink) and the vascular identity of blood vessels located in the subarachnoid space (cisterna interpendicularis, delimited by square in a , b ) was determined. d , e Consecutive sections show that AQP1int + /NKCC1 + cells are present in the smooth muscle cell layer of arterioles (arrowheads) and in the endothelium of capillaries and venules, respectively (curved arrowheads). f , g Vascular endothelial cells were labeled by lectin (WGA-FITC, green), followed by standard Immunolabeling. DAPI counterstain (blue) reveal the location of the leptomeningeal vessel (asterisk). h – j Higher magnification confocal images show that AQP1 is restricted to tunica media, where AQP1ext + smooth muscle cells, identified by their round soma (arrowheads) are observed, whereas AQP1 is not present in the endothelial cell layer (curved arrowheads). The arrow indicates a leptomeningeal cell, also AQP1ext + . BS, brain stem; Cb, cerebellum; cp, cerebral peduncle; Ctx, cerebral cortex; Hip, hippocampus; Hyp, hypothalamus; OB, olfactory bulb; Pn, pontine nuclei. Scale bars: a , b 2 mm; c – e 100 μm; f – j 50 μm
    Figure Legend Snippet: AQP1 and NKCC1 are present in smooth muscle and endothelial cells of the leptomeningeal vasculature. a , b Paraffin sections of adult mouse brain (P90) immunolabeled with anti-AQP1int or anti-NKCC1 (both brown). c Some sections were stained with hematoxylin (HE, pink) and the vascular identity of blood vessels located in the subarachnoid space (cisterna interpendicularis, delimited by square in a , b ) was determined. d , e Consecutive sections show that AQP1int + /NKCC1 + cells are present in the smooth muscle cell layer of arterioles (arrowheads) and in the endothelium of capillaries and venules, respectively (curved arrowheads). f , g Vascular endothelial cells were labeled by lectin (WGA-FITC, green), followed by standard Immunolabeling. DAPI counterstain (blue) reveal the location of the leptomeningeal vessel (asterisk). h – j Higher magnification confocal images show that AQP1 is restricted to tunica media, where AQP1ext + smooth muscle cells, identified by their round soma (arrowheads) are observed, whereas AQP1 is not present in the endothelial cell layer (curved arrowheads). The arrow indicates a leptomeningeal cell, also AQP1ext + . BS, brain stem; Cb, cerebellum; cp, cerebral peduncle; Ctx, cerebral cortex; Hip, hippocampus; Hyp, hypothalamus; OB, olfactory bulb; Pn, pontine nuclei. Scale bars: a , b 2 mm; c – e 100 μm; f – j 50 μm

    Techniques Used: Immunolabeling, Staining, Labeling, Whole Genome Amplification

    AQP1 and NKCC1 are expressed by the choroidal epithelial cells and in the leptomeningeal vasculature. a – f Confocal micrograph show a leptomeningeal WGA-FITC + (green) labeled vessel immunoreactive for AQP1 (magenta) and NKCC1 (orange) in the adult mouse brain (P90). In b an optical section reveals that AQP1 + /NKCC1 + cells are restricted to the smooth muscle cell layer (arrowheads) and absent in the endothelial cells (curved arrowheads), which are labeled by WGA-FITC. g , h NKCC1 is detected in the choroid plexus epithelia, in ependymal cells and in the molecular layer of the cerebellum, as shown in the micrographs of the fourth ventricle. i Double labeling confirms AQP1 and NKCC1 presence in choroid plexus epithelial cells (higher magnification of the area delimited by the blue square in h ). j , k Brain sections obtained from NKCC1 KO adult mice show no immunoreactivity in the brain parenchyma neither in the choroid plexus. l , m Histological sections immunolabeled with antibodies against AQP1ext (magenta), NKCC1 (yellow) and α-SMA (cyan), reveal AQP1ext + /NKCC1 + /α-SMA + leptomeningeal vessels around the hippocampus and third ventricle. Low magnification micrograph shows DAPI (blue) counterstaining and indicates a leptomeningeal blood vessel (asterisk) closely located to the hippocampal fissure. n – p Higher magnification of an AQP1ext + /NKCC1 + vessel (delimited by the dashed square in j . Arrowheads indicate α-SMA + cells. ( q ) Optical sectioning reveals that both AQP1 and NKCC1 are distributed in the smooth muscle cell layer (arrowheads). r 3D rendering of the leptomeningeal vessel confirms AQP1 and NKCC1 restriction to the smooth muscle cell layer (arrowheads). ChP, choroid plexus; DG, dentate gyrus; DS, dorsal subiculum; GL, granular layer; hif, hippocampal fissure; Mol, molecular layer; SAS, subarachnoid space; 3V, third ventricle, 4V, fourth ventricle. Scale bars: a , i 20 µm; b – f , q , r 10 µm; g , h , j – p 50 µm
    Figure Legend Snippet: AQP1 and NKCC1 are expressed by the choroidal epithelial cells and in the leptomeningeal vasculature. a – f Confocal micrograph show a leptomeningeal WGA-FITC + (green) labeled vessel immunoreactive for AQP1 (magenta) and NKCC1 (orange) in the adult mouse brain (P90). In b an optical section reveals that AQP1 + /NKCC1 + cells are restricted to the smooth muscle cell layer (arrowheads) and absent in the endothelial cells (curved arrowheads), which are labeled by WGA-FITC. g , h NKCC1 is detected in the choroid plexus epithelia, in ependymal cells and in the molecular layer of the cerebellum, as shown in the micrographs of the fourth ventricle. i Double labeling confirms AQP1 and NKCC1 presence in choroid plexus epithelial cells (higher magnification of the area delimited by the blue square in h ). j , k Brain sections obtained from NKCC1 KO adult mice show no immunoreactivity in the brain parenchyma neither in the choroid plexus. l , m Histological sections immunolabeled with antibodies against AQP1ext (magenta), NKCC1 (yellow) and α-SMA (cyan), reveal AQP1ext + /NKCC1 + /α-SMA + leptomeningeal vessels around the hippocampus and third ventricle. Low magnification micrograph shows DAPI (blue) counterstaining and indicates a leptomeningeal blood vessel (asterisk) closely located to the hippocampal fissure. n – p Higher magnification of an AQP1ext + /NKCC1 + vessel (delimited by the dashed square in j . Arrowheads indicate α-SMA + cells. ( q ) Optical sectioning reveals that both AQP1 and NKCC1 are distributed in the smooth muscle cell layer (arrowheads). r 3D rendering of the leptomeningeal vessel confirms AQP1 and NKCC1 restriction to the smooth muscle cell layer (arrowheads). ChP, choroid plexus; DG, dentate gyrus; DS, dorsal subiculum; GL, granular layer; hif, hippocampal fissure; Mol, molecular layer; SAS, subarachnoid space; 3V, third ventricle, 4V, fourth ventricle. Scale bars: a , i 20 µm; b – f , q , r 10 µm; g , h , j – p 50 µm

    Techniques Used: Whole Genome Amplification, Labeling, Mouse Assay, Immunolabeling

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    Alomone Labs anti aqp3
    <t>AQP3</t> Immunoreactivity was Mislocalized in a Majority of Psoriatic Lesions (A through C) Archived paraffin-embedded psoriasis samples were sectioned (4 μm) and deparaffinized. Sections were then stained with an antibody recognizing AQP3 and visualized with an ABC kit and DAB. Results from three patients are shown and the staining pattern is representative of 8 of 10 psoriatic samples examined. (D) A negative control was performed by omitting primary antibody. Note that AQP3 is localized to the cytoplasm rather than to the plasma membrane in a majority of the psoriatic epidermis. All sections were counterstained with hematoxylin.
    Anti Aqp3, 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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    Alomone Labs anti aqp5
    Expression of known AT1 cell markers in rat microarray data. Log2 expression data were generated from microarray experiments: AT1-like cells differentiated in culture ( blue ) (Day 2-6), freshly isolated AT1 cells ( purple ), freshly isolated AT2 cells ( red ), and other tissues ( black ). Table at bottom right includes previously described AT1 cell–specific genes, their associated Illumina Probe IDs, and their FDR-corrected P values in this study. FDR adjustment is based on the number of tests shown for known genes. AGER, advanced glycosylation end product–specific receptor; <t>AQP5,</t> aquaporin 5; CAV, caveolin; FDR, false-discovery rate; ILMN, Illumina probe number; PDPN, podoplanin. **Indicates significantly greater in rat AT1 and AT1-like cells compared to all others.
    Anti Aqp5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti aqp4
    Increased renal abundance of AQP2 but not <t>AQP4</t> in TRPC3 -/- mice. (A) Representative Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation probed with anti-AQP2 and anti β-actin antibodies. AQP2 appears as a duplet of the upper glycosylated near 37 kDa and lower non-glycosylated near 29 kDa bands. Each kidney was from separate animal. ( B ) Summary graph comparing total renal AQP2 expression (both glycosylated and non-glycosylated forms) in WT and TRPC3 -/- mice from Western blots similar to that shown in (A) Intensities of AQP2-reporting bands were normalized to the intensities of the respective actin bands. *—significant increase versus respective WT values. (C) Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation (indicated by a line) probed with anti-AQP4 antibodies. The Ponceau red staining of the same nitrocellulose membrane demonstrating equal protein loading is shown below. (D) Summary graph comparing total renal AQP4 expression for WT and TRPC3 -/- mice under tested conditions. The intensity values were normalized to the total signal of the respective lines in Ponceau red staining.
    Anti Aqp4, 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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    AQP3 Immunoreactivity was Mislocalized in a Majority of Psoriatic Lesions (A through C) Archived paraffin-embedded psoriasis samples were sectioned (4 μm) and deparaffinized. Sections were then stained with an antibody recognizing AQP3 and visualized with an ABC kit and DAB. Results from three patients are shown and the staining pattern is representative of 8 of 10 psoriatic samples examined. (D) A negative control was performed by omitting primary antibody. Note that AQP3 is localized to the cytoplasm rather than to the plasma membrane in a majority of the psoriatic epidermis. All sections were counterstained with hematoxylin.

    Journal: Archives of dermatological research

    Article Title: ABNORMAL AQUAPORIN-3 PROTEIN EXPRESSION IN HYPERPROLIFERATIVE SKIN DISORDERS

    doi: 10.1007/s00403-011-1136-x

    Figure Lengend Snippet: AQP3 Immunoreactivity was Mislocalized in a Majority of Psoriatic Lesions (A through C) Archived paraffin-embedded psoriasis samples were sectioned (4 μm) and deparaffinized. Sections were then stained with an antibody recognizing AQP3 and visualized with an ABC kit and DAB. Results from three patients are shown and the staining pattern is representative of 8 of 10 psoriatic samples examined. (D) A negative control was performed by omitting primary antibody. Note that AQP3 is localized to the cytoplasm rather than to the plasma membrane in a majority of the psoriatic epidermis. All sections were counterstained with hematoxylin.

    Article Snippet: Slides for AQP3 staining were deparaffinized, washed twice for 5 minutes in phosphate-buffered saline (PBS), incubated 30 minutes in 3% hydrogen peroxide, washed twice for 5 minutes in PBS, incubated 1 hour in 0.3% goat serum, and then incubated overnight with anti-AQP3 (Alomone Labs, Jerusalem, Israel; 1:1000 dilution) in a humidified chamber at 4°C.

    Techniques: Staining, Negative Control

    AQP3 Immunoreactivity was “Patchy” in Squamous Cell Carcinoma (SCC) Archived paraffin-embedded squamous cell carcinoma (SCC) samples were sectioned (4 μm) and deparaffinized. Sections were then stained with antibodies recognizing AQP3 and visualized with an ABC kit and DAB. Sections were counterstained with hematoxylin. Illustrated in panels A through C is AQP3 staining in 3 patients with results representative of 5 of 5 SCCs. A negative control in which the primary antibody was omitted showed no AQP3 staining (panel D).

    Journal: Archives of dermatological research

    Article Title: ABNORMAL AQUAPORIN-3 PROTEIN EXPRESSION IN HYPERPROLIFERATIVE SKIN DISORDERS

    doi: 10.1007/s00403-011-1136-x

    Figure Lengend Snippet: AQP3 Immunoreactivity was “Patchy” in Squamous Cell Carcinoma (SCC) Archived paraffin-embedded squamous cell carcinoma (SCC) samples were sectioned (4 μm) and deparaffinized. Sections were then stained with antibodies recognizing AQP3 and visualized with an ABC kit and DAB. Sections were counterstained with hematoxylin. Illustrated in panels A through C is AQP3 staining in 3 patients with results representative of 5 of 5 SCCs. A negative control in which the primary antibody was omitted showed no AQP3 staining (panel D).

    Article Snippet: Slides for AQP3 staining were deparaffinized, washed twice for 5 minutes in phosphate-buffered saline (PBS), incubated 30 minutes in 3% hydrogen peroxide, washed twice for 5 minutes in PBS, incubated 1 hour in 0.3% goat serum, and then incubated overnight with anti-AQP3 (Alomone Labs, Jerusalem, Israel; 1:1000 dilution) in a humidified chamber at 4°C.

    Techniques: Staining, Negative Control

    AQP3 Staining was Reduced in Ki67-Positive Cells in Squamous Cell Carcinoma Archived paraffin-embedded squamous cell carcinoma (SCC) samples were sectioned (4 μm) and deparaffinized. Sequential serial sections were then stained with antibodies recognizing (A, C) AQP3 or (B, D) Ki67 [an antigen expressed in proliferating cells (Dako, Carpinteria, CA)] and visualized with an ABC kit and DAB. Sections were counterstained with hematoxylin. Note that cells that are Ki67 positive exhibit reduced or absent AQP3 staining (arrows in panels C and D). The inverse correlation of Ki67 positivity with AQP3 immunoreactivity is representative of 4 of 4 SCCs. (E) Human tonsil stained with Ki67 as a positive control. (F) Primary antibody was omitted as a negative control.

    Journal: Archives of dermatological research

    Article Title: ABNORMAL AQUAPORIN-3 PROTEIN EXPRESSION IN HYPERPROLIFERATIVE SKIN DISORDERS

    doi: 10.1007/s00403-011-1136-x

    Figure Lengend Snippet: AQP3 Staining was Reduced in Ki67-Positive Cells in Squamous Cell Carcinoma Archived paraffin-embedded squamous cell carcinoma (SCC) samples were sectioned (4 μm) and deparaffinized. Sequential serial sections were then stained with antibodies recognizing (A, C) AQP3 or (B, D) Ki67 [an antigen expressed in proliferating cells (Dako, Carpinteria, CA)] and visualized with an ABC kit and DAB. Sections were counterstained with hematoxylin. Note that cells that are Ki67 positive exhibit reduced or absent AQP3 staining (arrows in panels C and D). The inverse correlation of Ki67 positivity with AQP3 immunoreactivity is representative of 4 of 4 SCCs. (E) Human tonsil stained with Ki67 as a positive control. (F) Primary antibody was omitted as a negative control.

    Article Snippet: Slides for AQP3 staining were deparaffinized, washed twice for 5 minutes in phosphate-buffered saline (PBS), incubated 30 minutes in 3% hydrogen peroxide, washed twice for 5 minutes in PBS, incubated 1 hour in 0.3% goat serum, and then incubated overnight with anti-AQP3 (Alomone Labs, Jerusalem, Israel; 1:1000 dilution) in a humidified chamber at 4°C.

    Techniques: Staining, Positive Control, Negative Control

    AQP3 Immunoreactivity was Reduced or Absent in Basal Cell Carcinoma (BCC) Archived paraffin-embedded basal cell carcinoma samples were sectioned (4 μm), deparaffinized, stained with antibodies recognizing AQP3 and visualized with an ABC kit and 3,3’-diaminobenzidine (DAB). Sections were counterstained with hematoxylin. Results illustrate three BCCs from different patients and are representative of 13 of 13 BCCs. The results were quantified as 3+ (intense staining), 2+ (moderate staining), + (weak staining) or 0 (no staining) by three independent observers and the values averaged. Shown in panel D is the quantitation (means ± SEM) of the 13 BCCs relative to normal-appearing overlying epidermis with the data analyzed using a student's t-test; *p

    Journal: Archives of dermatological research

    Article Title: ABNORMAL AQUAPORIN-3 PROTEIN EXPRESSION IN HYPERPROLIFERATIVE SKIN DISORDERS

    doi: 10.1007/s00403-011-1136-x

    Figure Lengend Snippet: AQP3 Immunoreactivity was Reduced or Absent in Basal Cell Carcinoma (BCC) Archived paraffin-embedded basal cell carcinoma samples were sectioned (4 μm), deparaffinized, stained with antibodies recognizing AQP3 and visualized with an ABC kit and 3,3’-diaminobenzidine (DAB). Sections were counterstained with hematoxylin. Results illustrate three BCCs from different patients and are representative of 13 of 13 BCCs. The results were quantified as 3+ (intense staining), 2+ (moderate staining), + (weak staining) or 0 (no staining) by three independent observers and the values averaged. Shown in panel D is the quantitation (means ± SEM) of the 13 BCCs relative to normal-appearing overlying epidermis with the data analyzed using a student's t-test; *p

    Article Snippet: Slides for AQP3 staining were deparaffinized, washed twice for 5 minutes in phosphate-buffered saline (PBS), incubated 30 minutes in 3% hydrogen peroxide, washed twice for 5 minutes in PBS, incubated 1 hour in 0.3% goat serum, and then incubated overnight with anti-AQP3 (Alomone Labs, Jerusalem, Israel; 1:1000 dilution) in a humidified chamber at 4°C.

    Techniques: Staining, Quantitation Assay

    Expression of known AT1 cell markers in rat microarray data. Log2 expression data were generated from microarray experiments: AT1-like cells differentiated in culture ( blue ) (Day 2-6), freshly isolated AT1 cells ( purple ), freshly isolated AT2 cells ( red ), and other tissues ( black ). Table at bottom right includes previously described AT1 cell–specific genes, their associated Illumina Probe IDs, and their FDR-corrected P values in this study. FDR adjustment is based on the number of tests shown for known genes. AGER, advanced glycosylation end product–specific receptor; AQP5, aquaporin 5; CAV, caveolin; FDR, false-discovery rate; ILMN, Illumina probe number; PDPN, podoplanin. **Indicates significantly greater in rat AT1 and AT1-like cells compared to all others.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Cross-Species Transcriptome Profiling Identifies New Alveolar Epithelial Type I Cell–Specific Genes

    doi: 10.1165/rcmb.2016-0071OC

    Figure Lengend Snippet: Expression of known AT1 cell markers in rat microarray data. Log2 expression data were generated from microarray experiments: AT1-like cells differentiated in culture ( blue ) (Day 2-6), freshly isolated AT1 cells ( purple ), freshly isolated AT2 cells ( red ), and other tissues ( black ). Table at bottom right includes previously described AT1 cell–specific genes, their associated Illumina Probe IDs, and their FDR-corrected P values in this study. FDR adjustment is based on the number of tests shown for known genes. AGER, advanced glycosylation end product–specific receptor; AQP5, aquaporin 5; CAV, caveolin; FDR, false-discovery rate; ILMN, Illumina probe number; PDPN, podoplanin. **Indicates significantly greater in rat AT1 and AT1-like cells compared to all others.

    Article Snippet: Blots were incubated with rabbit anti–epithelial sodium channel (ENaC) γ (1:200, sc-21014; Santa Cruz Biotechnology, Santa Cruz, CA), anti-semaphorin 3B (SEMA3B) (1:500, ; Abnova, Jhongli, Taiwan), anti-SEMA3E (1:100, AP7976b; Abgent, San Diego, CA), anti-GRAMD2 (1:100, ab84567; Abcam, Cambridge, MA), anti-SFTPC (1:200, AB3786; Millipore, Billerica, MA), and anti-AQP5 (1:200, AQP-005; Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Microarray, Generated, Isolation

    Validation of GRAMD2 expression in AT1 cells in lung tissue. ( A ) Confocal images for GRAMD2/SFTPC double staining in mouse lung sections show that GRAMD2 does not colocalize with SFTPC. DAPI is the nuclear counterstain. Scale bar : 20 µm. ( B ) Confocal images for GRAMD2/AQP5 double staining in mouse lung sections shows that GRAMD2 colocalizes with AQP5. DAPI is the nuclear counterstain. Scale bar : 20 µm. DAPI, 4’,6-diamidino-2-phenylindole.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Cross-Species Transcriptome Profiling Identifies New Alveolar Epithelial Type I Cell–Specific Genes

    doi: 10.1165/rcmb.2016-0071OC

    Figure Lengend Snippet: Validation of GRAMD2 expression in AT1 cells in lung tissue. ( A ) Confocal images for GRAMD2/SFTPC double staining in mouse lung sections show that GRAMD2 does not colocalize with SFTPC. DAPI is the nuclear counterstain. Scale bar : 20 µm. ( B ) Confocal images for GRAMD2/AQP5 double staining in mouse lung sections shows that GRAMD2 colocalizes with AQP5. DAPI is the nuclear counterstain. Scale bar : 20 µm. DAPI, 4’,6-diamidino-2-phenylindole.

    Article Snippet: Blots were incubated with rabbit anti–epithelial sodium channel (ENaC) γ (1:200, sc-21014; Santa Cruz Biotechnology, Santa Cruz, CA), anti-semaphorin 3B (SEMA3B) (1:500, ; Abnova, Jhongli, Taiwan), anti-SEMA3E (1:100, AP7976b; Abgent, San Diego, CA), anti-GRAMD2 (1:100, ab84567; Abcam, Cambridge, MA), anti-SFTPC (1:200, AB3786; Millipore, Billerica, MA), and anti-AQP5 (1:200, AQP-005; Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Double Staining

    Innervated salisphere-derived branching epithelial structures resemble an ex vivo fetal gland. D4 recombined mouse adult salispheres (A) and fetal epithelium (B) in laminin hydrogels with Mes, PSG and NRTN were stained for Peanut Agglutinin (PNA, red) to outline the epithelia, TUBB3 (green) for nerves, K5 (cyan), KIT (cyan), AQP5 (red), KI67 (green) for proliferation and/or DAPI (blue) for nuclei. Confocal images of 10μm sections. Scale bars, 100 μm (A) and 50 μm (B) .

    Journal: Biomaterials

    Article Title: Neurturin-containing laminin matrices support innervated branching epithelium from adult epithelial salispheres

    doi: 10.1016/j.biomaterials.2019.119245

    Figure Lengend Snippet: Innervated salisphere-derived branching epithelial structures resemble an ex vivo fetal gland. D4 recombined mouse adult salispheres (A) and fetal epithelium (B) in laminin hydrogels with Mes, PSG and NRTN were stained for Peanut Agglutinin (PNA, red) to outline the epithelia, TUBB3 (green) for nerves, K5 (cyan), KIT (cyan), AQP5 (red), KI67 (green) for proliferation and/or DAPI (blue) for nuclei. Confocal images of 10μm sections. Scale bars, 100 μm (A) and 50 μm (B) .

    Article Snippet: Samples were blocked for 90 minutes with 10% donkey serum (Jackson Laboratories, ME), 1% BSA, and MOM IgG blocking-reagent (Vector Laboratories, CA) in 0.1% PBS-Tween-20, and incubated with primary-antibodies overnight at 4°C, including rat-anti-Kit (1:100, R & D Systems, MN), rabbit-anti-Keratin-5 (1:2000, Covance Research, NJ), rat-anti-Keratin-19 (1:300, Developmental Biology Hybridoma Bank, University of Iowa) ), rabbit-Aqp5 (1:200, Alomone Labs, Israel), mouse-anti-SMA (1:200, Sigma Aldrich, MO), mouse-anti-Ki67 (BD Biosciences), rabbit-anti-Keratin-14 (1:2000, Covance Research, NJ) and rabbit-anti-E-cadherin (1:100,Cell Signaling Technology, MA).

    Techniques: Derivative Assay, Ex Vivo, Staining

    Increased renal abundance of AQP2 but not AQP4 in TRPC3 -/- mice. (A) Representative Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation probed with anti-AQP2 and anti β-actin antibodies. AQP2 appears as a duplet of the upper glycosylated near 37 kDa and lower non-glycosylated near 29 kDa bands. Each kidney was from separate animal. ( B ) Summary graph comparing total renal AQP2 expression (both glycosylated and non-glycosylated forms) in WT and TRPC3 -/- mice from Western blots similar to that shown in (A) Intensities of AQP2-reporting bands were normalized to the intensities of the respective actin bands. *—significant increase versus respective WT values. (C) Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation (indicated by a line) probed with anti-AQP4 antibodies. The Ponceau red staining of the same nitrocellulose membrane demonstrating equal protein loading is shown below. (D) Summary graph comparing total renal AQP4 expression for WT and TRPC3 -/- mice under tested conditions. The intensity values were normalized to the total signal of the respective lines in Ponceau red staining.

    Journal: PLoS ONE

    Article Title: TRPC3 determines osmosensitive [Ca2+]i signaling in the collecting duct and contributes to urinary concentration

    doi: 10.1371/journal.pone.0226381

    Figure Lengend Snippet: Increased renal abundance of AQP2 but not AQP4 in TRPC3 -/- mice. (A) Representative Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation probed with anti-AQP2 and anti β-actin antibodies. AQP2 appears as a duplet of the upper glycosylated near 37 kDa and lower non-glycosylated near 29 kDa bands. Each kidney was from separate animal. ( B ) Summary graph comparing total renal AQP2 expression (both glycosylated and non-glycosylated forms) in WT and TRPC3 -/- mice from Western blots similar to that shown in (A) Intensities of AQP2-reporting bands were normalized to the intensities of the respective actin bands. *—significant increase versus respective WT values. (C) Western blot from whole kidney lysates of WT and TRPC3 -/- mice at the baseline and after 24h water deprivation (indicated by a line) probed with anti-AQP4 antibodies. The Ponceau red staining of the same nitrocellulose membrane demonstrating equal protein loading is shown below. (D) Summary graph comparing total renal AQP4 expression for WT and TRPC3 -/- mice under tested conditions. The intensity values were normalized to the total signal of the respective lines in Ponceau red staining.

    Article Snippet: The primary antibodies were anti-AQP2 (1:1500, Alomone Labs, Israel; Cat. # AQP2-002), anti-AQP4 (1:1000, Alomone Labs, Israel; Cat. # AQP-004) and anti-β-actin (1:5000, Abcam, UK; Cat. # ab8227).

    Techniques: Mouse Assay, Western Blot, Expressing, Staining