kir4 1  (Alomone Labs)


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    Alomone Labs kir4 1
    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and <t>Kir4.1</t> + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.
    Kir4 1, 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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    Images

    1) Product Images from "Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes"

    Article Title: Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes

    Journal: Molecular Brain

    doi: 10.1186/1756-6606-6-28

    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.
    Figure Legend Snippet: Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.

    Techniques Used: Injection, Labeling, Software

    Primary antibodies used in immunohistochemistry
    Figure Legend Snippet: Primary antibodies used in immunohistochemistry

    Techniques Used:

    kir4 1  (Alomone Labs)


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    Alomone Labs kir4 1
    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and <t>Kir4.1</t> + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.
    Kir4 1, 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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    Images

    1) Product Images from "Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes"

    Article Title: Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes

    Journal: Molecular Brain

    doi: 10.1186/1756-6606-6-28

    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.
    Figure Legend Snippet: Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.

    Techniques Used: Injection, Labeling, Software

    Primary antibodies used in immunohistochemistry
    Figure Legend Snippet: Primary antibodies used in immunohistochemistry

    Techniques Used:

    kir4 1  (Alomone Labs)


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    Alomone Labs kir4 1
    The slices were derived from control and detached retina of C57BL/6 mice 24 h after surgery. A,G: Retinal sections were probed with antibodies against GFAP (green) and for nuclei (Dapi, blue). Note the upregulation of GFAP after RD. B,H: Sections were also stained with antibodies against <t>Kir4.1</t> (green). RD induced a mislocation of Kir4.1 along Müller cells (filled arrowhead) while the staining at the ILM (open arrowhead) and around blood vessels (arrow) remain unchanged. C,I: AQP4 staining (green) at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead) was reduced in detached retina. D,J: Retinal sections were probed with a pan specific antibody against all forms of dystrophins (green). Dystrophin-Dp71 staining is localized at the ILM (open arrowhead) and around blood vessels (arrow), as previously reported . RD induced a reduction of Dp71 staining. E,K: 24 h after surgery Immunoreactivity for Utrophin (green) at the ILM (open arrowhead) was upregulated after RD. In control retina utrophin was localized at the ILM, around blood vessels and in the OPL. F,L: Sections were stained with antibodies against β-dystroglycan (β-DG). In control retina, β-DG is localized at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead). RD induced a reduction of the staining in the ILM while the staining at the OPL (filled arrowhead) and around blood vessels (arrow) remain unchanged. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; ILM, inner limiting membrane. Scale bar = 30 µm.
    Kir4 1, 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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    Images

    1) Product Images from "Functional Implication of Dp71 in Osmoregulation and Vascular Permeability of the Retina"

    Article Title: Functional Implication of Dp71 in Osmoregulation and Vascular Permeability of the Retina

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007329

    The slices were derived from control and detached retina of C57BL/6 mice 24 h after surgery. A,G: Retinal sections were probed with antibodies against GFAP (green) and for nuclei (Dapi, blue). Note the upregulation of GFAP after RD. B,H: Sections were also stained with antibodies against Kir4.1 (green). RD induced a mislocation of Kir4.1 along Müller cells (filled arrowhead) while the staining at the ILM (open arrowhead) and around blood vessels (arrow) remain unchanged. C,I: AQP4 staining (green) at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead) was reduced in detached retina. D,J: Retinal sections were probed with a pan specific antibody against all forms of dystrophins (green). Dystrophin-Dp71 staining is localized at the ILM (open arrowhead) and around blood vessels (arrow), as previously reported . RD induced a reduction of Dp71 staining. E,K: 24 h after surgery Immunoreactivity for Utrophin (green) at the ILM (open arrowhead) was upregulated after RD. In control retina utrophin was localized at the ILM, around blood vessels and in the OPL. F,L: Sections were stained with antibodies against β-dystroglycan (β-DG). In control retina, β-DG is localized at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead). RD induced a reduction of the staining in the ILM while the staining at the OPL (filled arrowhead) and around blood vessels (arrow) remain unchanged. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; ILM, inner limiting membrane. Scale bar = 30 µm.
    Figure Legend Snippet: The slices were derived from control and detached retina of C57BL/6 mice 24 h after surgery. A,G: Retinal sections were probed with antibodies against GFAP (green) and for nuclei (Dapi, blue). Note the upregulation of GFAP after RD. B,H: Sections were also stained with antibodies against Kir4.1 (green). RD induced a mislocation of Kir4.1 along Müller cells (filled arrowhead) while the staining at the ILM (open arrowhead) and around blood vessels (arrow) remain unchanged. C,I: AQP4 staining (green) at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead) was reduced in detached retina. D,J: Retinal sections were probed with a pan specific antibody against all forms of dystrophins (green). Dystrophin-Dp71 staining is localized at the ILM (open arrowhead) and around blood vessels (arrow), as previously reported . RD induced a reduction of Dp71 staining. E,K: 24 h after surgery Immunoreactivity for Utrophin (green) at the ILM (open arrowhead) was upregulated after RD. In control retina utrophin was localized at the ILM, around blood vessels and in the OPL. F,L: Sections were stained with antibodies against β-dystroglycan (β-DG). In control retina, β-DG is localized at the ILM (open arrowhead), around blood vessels (arrow) and at the OPL (filled arrowhead). RD induced a reduction of the staining in the ILM while the staining at the OPL (filled arrowhead) and around blood vessels (arrow) remain unchanged. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; ILM, inner limiting membrane. Scale bar = 30 µm.

    Techniques Used: Derivative Assay, Staining

    A: 24 h after surgery, proteins of control and detached retina from C57BL/6 mice were extracted and Western blotting was performed. Representative photograph of immunoblots reacted with anti-GFAP, anti-Kir4.1, anti-AQP4, anti utrophins, anti β-dystroglycan (β-DG) and β-Actin antibodies. Numbers on the left refer to the relative electrophoretic mobility of prestained molecular mass standards in kiloDaltons. B: The relative protein level is expressed in arbitrary units. Each value represents the ratio of the specific band stain intensity normalized to β-Actin expression (TotalLab TL120, Nonlinear Inc, Durham NC, USA). In detached retina, GFAP expression level was significantly upregulated while AQP4 expression was downregulated. There was no significant difference in Kir4.1, utrophin and β-DG protein expression level after retinal detachment. All experiments were repeated at least three times, and the bars represent means + SE for triplicate data points; n = 4. *p<0.05; ***p<0.001 significant differences versus control.
    Figure Legend Snippet: A: 24 h after surgery, proteins of control and detached retina from C57BL/6 mice were extracted and Western blotting was performed. Representative photograph of immunoblots reacted with anti-GFAP, anti-Kir4.1, anti-AQP4, anti utrophins, anti β-dystroglycan (β-DG) and β-Actin antibodies. Numbers on the left refer to the relative electrophoretic mobility of prestained molecular mass standards in kiloDaltons. B: The relative protein level is expressed in arbitrary units. Each value represents the ratio of the specific band stain intensity normalized to β-Actin expression (TotalLab TL120, Nonlinear Inc, Durham NC, USA). In detached retina, GFAP expression level was significantly upregulated while AQP4 expression was downregulated. There was no significant difference in Kir4.1, utrophin and β-DG protein expression level after retinal detachment. All experiments were repeated at least three times, and the bars represent means + SE for triplicate data points; n = 4. *p<0.05; ***p<0.001 significant differences versus control.

    Techniques Used: Western Blot, Staining, Expressing

    kcnj10 antibodies  (Alomone Labs)


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    Alomone Labs kcnj10 antibodies
    AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). <t>Anti-KCNJ10</t> antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.
    Kcnj10 Antibodies, 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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    1) Product Images from "AAV8BP2 and AAV8 transduce the mammalian cochlear lateral wall and endolymphatic sac with high efficiency"

    Article Title: AAV8BP2 and AAV8 transduce the mammalian cochlear lateral wall and endolymphatic sac with high efficiency

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1016/j.omtm.2022.07.013

    AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). Anti-KCNJ10 antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.
    Figure Legend Snippet: AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). Anti-KCNJ10 antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.

    Techniques Used: Marker, Transduction

    apc 165  (Alomone Labs)


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    Alomone Labs apc 165
    Apc 165, 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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    kir 4 1  (Alomone Labs)


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    Alomone Labs kir 4 1

    Kir 4 1, 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
    https://www.bioz.com/result/kir 4 1/product/Alomone Labs
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    1) Product Images from "Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit"

    Article Title: Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit

    Journal: eLife

    doi: 10.7554/eLife.71379


    Figure Legend Snippet:

    Techniques Used: Sequencing, Western Blot, Immunofluorescence, Transduction

    antibody against kir4 1  (Alomone Labs)


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    Alomone Labs antibody against kir4 1
    Antibody Against Kir4 1, 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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    kir 4 1  (Alomone Labs)


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    Alomone Labs kir 4 1
    Primary antibodies used for immunohistochemistry (IHC) and immunofluorescence (IF)
    Kir 4 1, 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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    1) Product Images from "Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia"

    Article Title: Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.16701


    Figure Legend Snippet: Primary antibodies used for immunohistochemistry (IHC) and immunofluorescence (IF)

    Techniques Used: Immunohistochemistry, Immunofluorescence


    Figure Legend Snippet: Quantitative analysis of murine and canine spinal ganglia (SG)

    Techniques Used: Staining

    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video  ,  and Video  ,
    Figure Legend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video , and Video ,

    Techniques Used: Formalin-fixed Paraffin-Embedded, Marker, Staining

    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video  ,  and Video  ,
    Figure Legend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video , and Video ,

    Techniques Used: Formalin-fixed Paraffin-Embedded, Marker, Staining

    kir4 1  (Alomone Labs)


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    Alomone Labs kir4 1
    Deletion of Nedd4-2 stimulates <t>Kir4.1/Kir5.1</t> in the DCT. (A) Representative single-channel recordings showing the basolateral K+ channel activity in the DCT of WT and Ks-Nedd4-2 KO mice. The experiments were performed in cell-attached patches with bath solution containing 140 mM NaCl and 5 mM KCl and pipette solution containing 140 mM KCl. The channel closed level is indicated by “C,” and the holding potential was 0 mV. (B) Probability of finding basolateral K+ channel activity, mean NPo, and Po in the DCT of WT and Ks-Nedd4-2 KO mice. *Significant difference between WT and Ks-Nedd4-2 KO mice. (C) An immunoblot showing the expression of Kir4.1 and Nedd4-2 in WT and Ks-Nedd4-2 KO mice. The sample from Ks-Kir4.1 KO mice serves as a negative control for Kir4.1. IB, immunoblot.
    Kir4 1, 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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    Images

    1) Product Images from "Renal Tubule Nedd4-2 Deficiency Stimulates Kir4.1/Kir5.1 and Thiazide-Sensitive NaCl Cotransporter in Distal Convoluted Tubule"

    Article Title: Renal Tubule Nedd4-2 Deficiency Stimulates Kir4.1/Kir5.1 and Thiazide-Sensitive NaCl Cotransporter in Distal Convoluted Tubule

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2019090923

    Deletion of Nedd4-2 stimulates Kir4.1/Kir5.1 in the DCT. (A) Representative single-channel recordings showing the basolateral K+ channel activity in the DCT of WT and Ks-Nedd4-2 KO mice. The experiments were performed in cell-attached patches with bath solution containing 140 mM NaCl and 5 mM KCl and pipette solution containing 140 mM KCl. The channel closed level is indicated by “C,” and the holding potential was 0 mV. (B) Probability of finding basolateral K+ channel activity, mean NPo, and Po in the DCT of WT and Ks-Nedd4-2 KO mice. *Significant difference between WT and Ks-Nedd4-2 KO mice. (C) An immunoblot showing the expression of Kir4.1 and Nedd4-2 in WT and Ks-Nedd4-2 KO mice. The sample from Ks-Kir4.1 KO mice serves as a negative control for Kir4.1. IB, immunoblot.
    Figure Legend Snippet: Deletion of Nedd4-2 stimulates Kir4.1/Kir5.1 in the DCT. (A) Representative single-channel recordings showing the basolateral K+ channel activity in the DCT of WT and Ks-Nedd4-2 KO mice. The experiments were performed in cell-attached patches with bath solution containing 140 mM NaCl and 5 mM KCl and pipette solution containing 140 mM KCl. The channel closed level is indicated by “C,” and the holding potential was 0 mV. (B) Probability of finding basolateral K+ channel activity, mean NPo, and Po in the DCT of WT and Ks-Nedd4-2 KO mice. *Significant difference between WT and Ks-Nedd4-2 KO mice. (C) An immunoblot showing the expression of Kir4.1 and Nedd4-2 in WT and Ks-Nedd4-2 KO mice. The sample from Ks-Kir4.1 KO mice serves as a negative control for Kir4.1. IB, immunoblot.

    Techniques Used: Activity Assay, Transferring, Western Blot, Expressing, Negative Control

    Double deletion of Nedd4-2 and Kir4.1 depolarizes DCT membrane. (A) Representative whole-cell recordings showing the Ba2+-sensitive K+ currents measured with ramp protocol from −100 to 100 mV in the DCT1 of WT mice (Kcnj10flox/flox/Nedd4lflox/flox) and Ks-Nedd4-2/Kir4.1 KO mice (dotted line). (B) A scatter graph summarizes the values measured at–60 mV with whole-cell recording. The mean value and SEM are shown on the left of each row. For the whole-cell recording, a symmetric 140 mM K+ solution was used for the bath and the pipette. (C) A perforated whole-cell recording showing the K+ current (IK) reversal potential in the DCT of WT and Ks-Nedd4-2/Kir4.1 KO mice. (D) A scatter graph summarizes the results of experiments in which the IK reversal potentials were measured in WT and Ks-Nedd4-2/Kir4.1 KO mice. The mean value and SEM are shown on the left of each row. For the measurement of IK reversal potential, the bath solution contains 140 mM NaCl and 5 mM KCl, whereas the pipette solution has 140 mM KCl. Data for WT (Nedd4lf/f) and Ks-Nedd4-2 KO mice in (B) and (D) are duplicated from Figure 2, B and D, respectively, for the comparison. *Significant difference (P=0.05) determined by t test.
    Figure Legend Snippet: Double deletion of Nedd4-2 and Kir4.1 depolarizes DCT membrane. (A) Representative whole-cell recordings showing the Ba2+-sensitive K+ currents measured with ramp protocol from −100 to 100 mV in the DCT1 of WT mice (Kcnj10flox/flox/Nedd4lflox/flox) and Ks-Nedd4-2/Kir4.1 KO mice (dotted line). (B) A scatter graph summarizes the values measured at–60 mV with whole-cell recording. The mean value and SEM are shown on the left of each row. For the whole-cell recording, a symmetric 140 mM K+ solution was used for the bath and the pipette. (C) A perforated whole-cell recording showing the K+ current (IK) reversal potential in the DCT of WT and Ks-Nedd4-2/Kir4.1 KO mice. (D) A scatter graph summarizes the results of experiments in which the IK reversal potentials were measured in WT and Ks-Nedd4-2/Kir4.1 KO mice. The mean value and SEM are shown on the left of each row. For the measurement of IK reversal potential, the bath solution contains 140 mM NaCl and 5 mM KCl, whereas the pipette solution has 140 mM KCl. Data for WT (Nedd4lf/f) and Ks-Nedd4-2 KO mice in (B) and (D) are duplicated from Figure 2, B and D, respectively, for the comparison. *Significant difference (P=0.05) determined by t test.

    Techniques Used: Transferring

    Deletion of Nedd4-2 increases the expression of ENaCα. (A) An immunoblot showing the expression of full-length (f) ENaCα and cleaved (c) ENaCα in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Nedd4lflox/flox-Kcnj10flox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice. Gapdh, glyceraldehyde-3-phosphate dehydrogenase; IB, immunoblot. A scatter graph summarizes the normalized band intensity of the above experiments for (B) f-ENaCα and (C) c-ENaCα. The mean value and SEM are shown on the left of each row. *Difference is significant at P=0.05; **difference is significant at P=0.01.
    Figure Legend Snippet: Deletion of Nedd4-2 increases the expression of ENaCα. (A) An immunoblot showing the expression of full-length (f) ENaCα and cleaved (c) ENaCα in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Nedd4lflox/flox-Kcnj10flox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice. Gapdh, glyceraldehyde-3-phosphate dehydrogenase; IB, immunoblot. A scatter graph summarizes the normalized band intensity of the above experiments for (B) f-ENaCα and (C) c-ENaCα. The mean value and SEM are shown on the left of each row. *Difference is significant at P=0.05; **difference is significant at P=0.01.

    Techniques Used: Expressing, Western Blot

    NCC expression is not increased in the double-KO mice. (A) An immunoblot showing the expression of pNCC, tNCC, Kir4.1, and Nedd4-2 in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice. Gapdh, glyceraldehyde-3-phosphate dehydrogenase; IB, immunoblot. A scatter graph summarizes the normalized band intensity of the (B) pNCC and (C) tNCC in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice, respectively. The mean value and SEM are shown on the left of each row. Both male and female mice were used for the western blot, and the data obtained from male/female mice were normalized with corresponding male/female WT mice. *Significant difference (P=0.05) determined by t test; #value is significantly different between Ks-Kir4.1 KO and Ks-Nedd4-2/Kir4.1 KO mice.
    Figure Legend Snippet: NCC expression is not increased in the double-KO mice. (A) An immunoblot showing the expression of pNCC, tNCC, Kir4.1, and Nedd4-2 in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice. Gapdh, glyceraldehyde-3-phosphate dehydrogenase; IB, immunoblot. A scatter graph summarizes the normalized band intensity of the (B) pNCC and (C) tNCC in WT (Kcnj10flox/flox, Nedd4lflox/flox, and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, Ks-Nedd4-2 KO, and Ks-Nedd4-2/Kir4.1 KO mice, respectively. The mean value and SEM are shown on the left of each row. Both male and female mice were used for the western blot, and the data obtained from male/female mice were normalized with corresponding male/female WT mice. *Significant difference (P=0.05) determined by t test; #value is significantly different between Ks-Kir4.1 KO and Ks-Nedd4-2/Kir4.1 KO mice.

    Techniques Used: Expressing, Western Blot

    Nedd4-2 deletion delays Kir4.1 deletion-induced inhibition of NCC. (A) An immunoblot showing the expression of pNCC, tNCC, Kir4.1, and Nedd4-2 in WT (Kcnj10flox/flox and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, and Ks-Nedd4-2/Kir4.1 KO mice. The experiments was conducted in the mice immediately after treatment of doxycycline without a 14-day transition period. Gapdh, glyceraldehyde-3-phosphate dehydrogenase. (B) A scatter graph summarizes the normalized band intensity of the pNCC (left panel) and tNCC (right panel) in WT (Kcnj10flox/flox and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, and Ks-Nedd4-2/Kir4.1 KO mice. The mean value and SEM are shown on the left of each row. Male mice were used for the western blot. (C) A scatter plot shows the HCTZ-induced net renal Na+ excretion in WT, Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. The mice were used immediately after either doxycycline or vehicle treatment. BW, body weight.
    Figure Legend Snippet: Nedd4-2 deletion delays Kir4.1 deletion-induced inhibition of NCC. (A) An immunoblot showing the expression of pNCC, tNCC, Kir4.1, and Nedd4-2 in WT (Kcnj10flox/flox and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, and Ks-Nedd4-2/Kir4.1 KO mice. The experiments was conducted in the mice immediately after treatment of doxycycline without a 14-day transition period. Gapdh, glyceraldehyde-3-phosphate dehydrogenase. (B) A scatter graph summarizes the normalized band intensity of the pNCC (left panel) and tNCC (right panel) in WT (Kcnj10flox/flox and Kcnj10flox/flox/Nedd4lflox/flox), Ks-Kir4.1 KO, and Ks-Nedd4-2/Kir4.1 KO mice. The mean value and SEM are shown on the left of each row. Male mice were used for the western blot. (C) A scatter plot shows the HCTZ-induced net renal Na+ excretion in WT, Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. The mice were used immediately after either doxycycline or vehicle treatment. BW, body weight.

    Techniques Used: Inhibition, Western Blot, Expressing

    NCC expression is suppressed in Ks-Nedd4-2/Kir4.1 KO mice. Images show tNCC immunostaining in (A–C) WT (Nedd4lflox/flox/Kcnj10flox/flox), (D–F) Ks-Nedd4-2/Kir4.1 KO, (G–I) Ks-Nedd4-2 KO, and (J–L) Ks-Kir4.1 KO mice. The images in red squares in (A, D, G, and J) are magnified with intermediate (B, E, H, and K) and large resolution (C, F, I, and L). Original magnification, ×4 in A, D, G, and J; ×20 in B, E, H, and K; ×60 in C, F, I, and L.
    Figure Legend Snippet: NCC expression is suppressed in Ks-Nedd4-2/Kir4.1 KO mice. Images show tNCC immunostaining in (A–C) WT (Nedd4lflox/flox/Kcnj10flox/flox), (D–F) Ks-Nedd4-2/Kir4.1 KO, (G–I) Ks-Nedd4-2 KO, and (J–L) Ks-Kir4.1 KO mice. The images in red squares in (A, D, G, and J) are magnified with intermediate (B, E, H, and K) and large resolution (C, F, I, and L). Original magnification, ×4 in A, D, G, and J; ×20 in B, E, H, and K; ×60 in C, F, I, and L.

    Techniques Used: Expressing, Immunostaining

    Double deletion of Kir4.1 and Nedd4-2 impairs NCC function. (A) A scatter plot showing the Δ values of HCTZ-induced net ENa in WT (Nedd4lflox/flox/Kcnj10flox/flox), Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. (B) A scatter line graph showing the results of each experiment in which the effect of single-dose HCTZ (30 mg/kg body wt [BW]) on urinary Na+ excretion (ENa) within 120 minutes was measured with the renal clearance method in WT (Nedd4lflox/flox/Kcnj10flox/flox) and Ks-Nedd4-2/Kir4.1 KO mice. The significance is determined by t test. (C) A scatter line graph showing the results of each experiment in which the effect of single-dose HCTZ (30 mg/kg BW) on urinary K+ excretion (EK) within 120 minutes was measured with the renal clearance method in WT (Nedd4lflox/flox/Kcnj10flox/flox) and Ks-Nedd4-2/Kir4.1 KO mice. (D) A scatter plot shows the plasma K+ concentrations in WT (Nedd4lflox/flox and Nedd4lflox/flox/Kcnj10flox/flox), Ks-Nedd4-2 KO, Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. The experiments were performed in the mice treated with doxycycline followed by a 14-day recovery period.
    Figure Legend Snippet: Double deletion of Kir4.1 and Nedd4-2 impairs NCC function. (A) A scatter plot showing the Δ values of HCTZ-induced net ENa in WT (Nedd4lflox/flox/Kcnj10flox/flox), Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. (B) A scatter line graph showing the results of each experiment in which the effect of single-dose HCTZ (30 mg/kg body wt [BW]) on urinary Na+ excretion (ENa) within 120 minutes was measured with the renal clearance method in WT (Nedd4lflox/flox/Kcnj10flox/flox) and Ks-Nedd4-2/Kir4.1 KO mice. The significance is determined by t test. (C) A scatter line graph showing the results of each experiment in which the effect of single-dose HCTZ (30 mg/kg BW) on urinary K+ excretion (EK) within 120 minutes was measured with the renal clearance method in WT (Nedd4lflox/flox/Kcnj10flox/flox) and Ks-Nedd4-2/Kir4.1 KO mice. (D) A scatter plot shows the plasma K+ concentrations in WT (Nedd4lflox/flox and Nedd4lflox/flox/Kcnj10flox/flox), Ks-Nedd4-2 KO, Ks-Nedd4-2/Kir4.1 KO, and Ks-Kir4.1 KO mice. The experiments were performed in the mice treated with doxycycline followed by a 14-day recovery period.

    Techniques Used:

    A cell scheme illustrates the role of Kir4.1/Kir5.1 in mediating the effect of Nedd4-2 on thiazide-sensitive NCC. The inhibition of Kir4.1/Kir5.1 activity by Nedd4-2 should depolarize the DCT membrane voltage (V). A decrease in the cell negativity reduces the driving force for Cl− exit by basolateral Cl− channels, thereby increasing intracellular Cl− (Cl−i), which inhibits WNK and SPAK. This leads to inhibition of NCC phosphorylation and activity. Nedd4-2 is expected to facilitate the degradation of non-pNCC. In addition, Nedd4-2 also regulates NCC retrieval from the apical membrane of the DCT by Kir4.1-indipendent mechanism (blue arrow). The dotted line indicates the inhibition.
    Figure Legend Snippet: A cell scheme illustrates the role of Kir4.1/Kir5.1 in mediating the effect of Nedd4-2 on thiazide-sensitive NCC. The inhibition of Kir4.1/Kir5.1 activity by Nedd4-2 should depolarize the DCT membrane voltage (V). A decrease in the cell negativity reduces the driving force for Cl− exit by basolateral Cl− channels, thereby increasing intracellular Cl− (Cl−i), which inhibits WNK and SPAK. This leads to inhibition of NCC phosphorylation and activity. Nedd4-2 is expected to facilitate the degradation of non-pNCC. In addition, Nedd4-2 also regulates NCC retrieval from the apical membrane of the DCT by Kir4.1-indipendent mechanism (blue arrow). The dotted line indicates the inhibition.

    Techniques Used: Inhibition, Activity Assay

    kir4 1  (Alomone Labs)


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    Structured Review

    Alomone Labs kir4 1
    Primers used for genotyping
    Kir4 1, 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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    Average 94 stars, based on 1 article reviews
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    Images

    1) Product Images from "Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron"

    Article Title: Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00059.2018

    Primers used for genotyping
    Figure Legend Snippet: Primers used for genotyping

    Techniques Used: Sequencing

    Kir 5.1 is required for Nedd4-2-mediated ubiquitination of Kir4.1. A: Ubiquitination assay shows ubiquitinated K channel in HEK293 cells transfected with Flag-tagged Kir4.1 or Kir5.1, Kir4.1+Nedd4-2, Kir4.1+Kir5.1, Kir5.1+Nedd4-2, and Kir4.1/5.1+ Nedd4-2, respectively. Flag antibody was used to immunoprecipitate Flag-tagged Kir4.1 or 5.1, and ubiquitin (Ub) antibody was used to detect ubiquitinated K channels (indicated by a bracket). B: Ubiquitination assay with SDS pretreatment shows ubiquitinated K channels in HEK293 cells transfected with Flag-tagged Kir4.1, Kir4.1+Nedd4-2, and Kir4.1/5.1+Nedd4-2. Lower panel shows the expression of Kir4.1 used for immunoprecipitation (IP). IB, immunoblot.
    Figure Legend Snippet: Kir 5.1 is required for Nedd4-2-mediated ubiquitination of Kir4.1. A: Ubiquitination assay shows ubiquitinated K channel in HEK293 cells transfected with Flag-tagged Kir4.1 or Kir5.1, Kir4.1+Nedd4-2, Kir4.1+Kir5.1, Kir5.1+Nedd4-2, and Kir4.1/5.1+ Nedd4-2, respectively. Flag antibody was used to immunoprecipitate Flag-tagged Kir4.1 or 5.1, and ubiquitin (Ub) antibody was used to detect ubiquitinated K channels (indicated by a bracket). B: Ubiquitination assay with SDS pretreatment shows ubiquitinated K channels in HEK293 cells transfected with Flag-tagged Kir4.1, Kir4.1+Nedd4-2, and Kir4.1/5.1+Nedd4-2. Lower panel shows the expression of Kir4.1 used for immunoprecipitation (IP). IB, immunoblot.

    Techniques Used: Ubiquitin Assay, Transfection, Expressing, Immunoprecipitation, Western Blot

    Nedd4-2 but not Nedd4-1 inhibits Kir4.1/Kir5.1 channels. Whole-cell recording shows Ba2+-sensitive K currents measured from −60 mV to 60 mV at 20-mV steps in HEK293 cells transfected with Kir4.1+Kir5.1 (A) or with Kir4.1/5.1 +Nedd4-2 (B). The symmetrical 145 mM K solution was used for both bath and pipette. C: Bar graph summarizes the results of experiments (n = 6) in which Ba2+-sensitive K currents were measured with whole-cell recording in HEK293 cells transfected with Kir4.1, Kir4.1+Nedd4-2, Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1+Nedd4-1, and Kir4.1/5.1+dead Nedd4-2. The patch-clamp experiments were performed 24 h after the transfection and the positive transfected cells were identified by green fluorescent protein (GFP) fluorescence. pA, picoamperes; pF, picofarads.
    Figure Legend Snippet: Nedd4-2 but not Nedd4-1 inhibits Kir4.1/Kir5.1 channels. Whole-cell recording shows Ba2+-sensitive K currents measured from −60 mV to 60 mV at 20-mV steps in HEK293 cells transfected with Kir4.1+Kir5.1 (A) or with Kir4.1/5.1 +Nedd4-2 (B). The symmetrical 145 mM K solution was used for both bath and pipette. C: Bar graph summarizes the results of experiments (n = 6) in which Ba2+-sensitive K currents were measured with whole-cell recording in HEK293 cells transfected with Kir4.1, Kir4.1+Nedd4-2, Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1+Nedd4-1, and Kir4.1/5.1+dead Nedd4-2. The patch-clamp experiments were performed 24 h after the transfection and the positive transfected cells were identified by green fluorescent protein (GFP) fluorescence. pA, picoamperes; pF, picofarads.

    Techniques Used: Transfection, Transferring, Patch Clamp, Fluorescence

    Mutation of TPVT motif of Kir5.1 abolishes Nedd4-2-mediated inhibition of Kir4.1/5.1. A: Western blot (IB) shows the expression of Kir4.1 in HEK293 cells transfected with Ki4.1+Nedd4-2, Kir4.1/5.1+Nedd4-2, Kir4.1+Nedd4-1, and Kir4.1/5.1+Nedd4-1. The expression of Nedd4 was shown in the middle panel. B: Normalized band density of Kir4.1 expression is shown in a bar graph. C: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents were measured (at −60 mV) with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1T249A+Nedd4-2, and Kir4.1/5.1T249D +Nedd4-2, respectively (n = 6). D: Ba2+-sensitive K currents measured at −60 mV with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1T249A, and Kir4.1/5.1T249D (n = 6). Patch-clamp experiments were performed 24 h after the transfection, and symmetrical 145 mM K was used for the pipette and the bath solution. pA, picoamperes; pF, picofarads.
    Figure Legend Snippet: Mutation of TPVT motif of Kir5.1 abolishes Nedd4-2-mediated inhibition of Kir4.1/5.1. A: Western blot (IB) shows the expression of Kir4.1 in HEK293 cells transfected with Ki4.1+Nedd4-2, Kir4.1/5.1+Nedd4-2, Kir4.1+Nedd4-1, and Kir4.1/5.1+Nedd4-1. The expression of Nedd4 was shown in the middle panel. B: Normalized band density of Kir4.1 expression is shown in a bar graph. C: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents were measured (at −60 mV) with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1T249A+Nedd4-2, and Kir4.1/5.1T249D +Nedd4-2, respectively (n = 6). D: Ba2+-sensitive K currents measured at −60 mV with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1T249A, and Kir4.1/5.1T249D (n = 6). Patch-clamp experiments were performed 24 h after the transfection, and symmetrical 145 mM K was used for the pipette and the bath solution. pA, picoamperes; pF, picofarads.

    Techniques Used: Mutagenesis, Inhibition, Western Blot, Expressing, Transfection, Patch Clamp, Transferring

    Deletion of Kir5.1 stimulates the basolateral Kir4.1 in the distal convoluted tubule (DCT). A: Whole-cell recording showing Ba2+-sensitive K currents in the DCT of the wild-type (WT) or Kir5.1 knockout (KO) mice. K currents were measured with a ramp protocol from −100 to 100 mV using symmetrical 145 mM K solution in the bath and pipette. B: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the DCT were measured at −60 mV from the WT and Kir5.1 KO mice (n = 6). Western blot (IB) shows the expression of Kir4.1 in WT and Kir5.1 KO mice (C), and a bar graph summarizes normalized band density of Kir4.1 (D). E: Immunostaining of Kir4.1 in kidney from WT and Kir5.1 KO mice. Treatment for the kidney slides was identical. pA, picoamperes.
    Figure Legend Snippet: Deletion of Kir5.1 stimulates the basolateral Kir4.1 in the distal convoluted tubule (DCT). A: Whole-cell recording showing Ba2+-sensitive K currents in the DCT of the wild-type (WT) or Kir5.1 knockout (KO) mice. K currents were measured with a ramp protocol from −100 to 100 mV using symmetrical 145 mM K solution in the bath and pipette. B: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the DCT were measured at −60 mV from the WT and Kir5.1 KO mice (n = 6). Western blot (IB) shows the expression of Kir4.1 in WT and Kir5.1 KO mice (C), and a bar graph summarizes normalized band density of Kir4.1 (D). E: Immunostaining of Kir4.1 in kidney from WT and Kir5.1 KO mice. Treatment for the kidney slides was identical. pA, picoamperes.

    Techniques Used: Knock-Out, Transferring, Western Blot, Expressing, Immunostaining

    Deletion of Nedd4-2 increases Kir4.1 expression. A: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the distal convoluted tubule (DCT) were measured at −60 mV from the wild-type (WT) and Ks-Nedd4-2 knockout (KO) mice (n = 6). B: Western blots (IB) showed the expression of Kir4.1 and Kir5.1 in WT and Ks-Nedd4-2 KO mice. A bar graph summarizes normalized band density of Kir4.1 and Ki5.1 (right panel). C: immunostaining of Kir4.1 in kidney from Ks-Nedd4-2 KO mice and WT control (without doxycycline treated). pA, picoamperes.
    Figure Legend Snippet: Deletion of Nedd4-2 increases Kir4.1 expression. A: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the distal convoluted tubule (DCT) were measured at −60 mV from the wild-type (WT) and Ks-Nedd4-2 knockout (KO) mice (n = 6). B: Western blots (IB) showed the expression of Kir4.1 and Kir5.1 in WT and Ks-Nedd4-2 KO mice. A bar graph summarizes normalized band density of Kir4.1 and Ki5.1 (right panel). C: immunostaining of Kir4.1 in kidney from Ks-Nedd4-2 KO mice and WT control (without doxycycline treated). pA, picoamperes.

    Techniques Used: Expressing, Knock-Out, Western Blot, Immunostaining

    A scheme illustrating the role of Kir5.1 as a binding partner in mediating Nedd4-2 E3 ligase dependent degradation of Kir4.1 in the distal convoluted tubule (DCT). A part of Kir5.1 sequence (from AA241 to 260) including TPVT motif is shown on the top. Ub, ubiquitin.
    Figure Legend Snippet: A scheme illustrating the role of Kir5.1 as a binding partner in mediating Nedd4-2 E3 ligase dependent degradation of Kir4.1 in the distal convoluted tubule (DCT). A part of Kir5.1 sequence (from AA241 to 260) including TPVT motif is shown on the top. Ub, ubiquitin.

    Techniques Used: Binding Assay, Sequencing

    kir4 1  (Alomone Labs)


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    Structured Review

    Alomone Labs kir4 1
    Primers used for genotyping
    Kir4 1, 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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    Images

    1) Product Images from "Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron"

    Article Title: Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00059.2018

    Primers used for genotyping
    Figure Legend Snippet: Primers used for genotyping

    Techniques Used: Sequencing

    Kir 5.1 is required for Nedd4-2-mediated ubiquitination of Kir4.1. A: Ubiquitination assay shows ubiquitinated K channel in HEK293 cells transfected with Flag-tagged Kir4.1 or Kir5.1, Kir4.1+Nedd4-2, Kir4.1+Kir5.1, Kir5.1+Nedd4-2, and Kir4.1/5.1+ Nedd4-2, respectively. Flag antibody was used to immunoprecipitate Flag-tagged Kir4.1 or 5.1, and ubiquitin (Ub) antibody was used to detect ubiquitinated K channels (indicated by a bracket). B: Ubiquitination assay with SDS pretreatment shows ubiquitinated K channels in HEK293 cells transfected with Flag-tagged Kir4.1, Kir4.1+Nedd4-2, and Kir4.1/5.1+Nedd4-2. Lower panel shows the expression of Kir4.1 used for immunoprecipitation (IP). IB, immunoblot.
    Figure Legend Snippet: Kir 5.1 is required for Nedd4-2-mediated ubiquitination of Kir4.1. A: Ubiquitination assay shows ubiquitinated K channel in HEK293 cells transfected with Flag-tagged Kir4.1 or Kir5.1, Kir4.1+Nedd4-2, Kir4.1+Kir5.1, Kir5.1+Nedd4-2, and Kir4.1/5.1+ Nedd4-2, respectively. Flag antibody was used to immunoprecipitate Flag-tagged Kir4.1 or 5.1, and ubiquitin (Ub) antibody was used to detect ubiquitinated K channels (indicated by a bracket). B: Ubiquitination assay with SDS pretreatment shows ubiquitinated K channels in HEK293 cells transfected with Flag-tagged Kir4.1, Kir4.1+Nedd4-2, and Kir4.1/5.1+Nedd4-2. Lower panel shows the expression of Kir4.1 used for immunoprecipitation (IP). IB, immunoblot.

    Techniques Used: Ubiquitin Assay, Transfection, Expressing, Immunoprecipitation, Western Blot

    Nedd4-2 but not Nedd4-1 inhibits Kir4.1/Kir5.1 channels. Whole-cell recording shows Ba2+-sensitive K currents measured from −60 mV to 60 mV at 20-mV steps in HEK293 cells transfected with Kir4.1+Kir5.1 (A) or with Kir4.1/5.1 +Nedd4-2 (B). The symmetrical 145 mM K solution was used for both bath and pipette. C: Bar graph summarizes the results of experiments (n = 6) in which Ba2+-sensitive K currents were measured with whole-cell recording in HEK293 cells transfected with Kir4.1, Kir4.1+Nedd4-2, Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1+Nedd4-1, and Kir4.1/5.1+dead Nedd4-2. The patch-clamp experiments were performed 24 h after the transfection and the positive transfected cells were identified by green fluorescent protein (GFP) fluorescence. pA, picoamperes; pF, picofarads.
    Figure Legend Snippet: Nedd4-2 but not Nedd4-1 inhibits Kir4.1/Kir5.1 channels. Whole-cell recording shows Ba2+-sensitive K currents measured from −60 mV to 60 mV at 20-mV steps in HEK293 cells transfected with Kir4.1+Kir5.1 (A) or with Kir4.1/5.1 +Nedd4-2 (B). The symmetrical 145 mM K solution was used for both bath and pipette. C: Bar graph summarizes the results of experiments (n = 6) in which Ba2+-sensitive K currents were measured with whole-cell recording in HEK293 cells transfected with Kir4.1, Kir4.1+Nedd4-2, Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1+Nedd4-1, and Kir4.1/5.1+dead Nedd4-2. The patch-clamp experiments were performed 24 h after the transfection and the positive transfected cells were identified by green fluorescent protein (GFP) fluorescence. pA, picoamperes; pF, picofarads.

    Techniques Used: Transfection, Transferring, Patch Clamp, Fluorescence

    Mutation of TPVT motif of Kir5.1 abolishes Nedd4-2-mediated inhibition of Kir4.1/5.1. A: Western blot (IB) shows the expression of Kir4.1 in HEK293 cells transfected with Ki4.1+Nedd4-2, Kir4.1/5.1+Nedd4-2, Kir4.1+Nedd4-1, and Kir4.1/5.1+Nedd4-1. The expression of Nedd4 was shown in the middle panel. B: Normalized band density of Kir4.1 expression is shown in a bar graph. C: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents were measured (at −60 mV) with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1T249A+Nedd4-2, and Kir4.1/5.1T249D +Nedd4-2, respectively (n = 6). D: Ba2+-sensitive K currents measured at −60 mV with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1T249A, and Kir4.1/5.1T249D (n = 6). Patch-clamp experiments were performed 24 h after the transfection, and symmetrical 145 mM K was used for the pipette and the bath solution. pA, picoamperes; pF, picofarads.
    Figure Legend Snippet: Mutation of TPVT motif of Kir5.1 abolishes Nedd4-2-mediated inhibition of Kir4.1/5.1. A: Western blot (IB) shows the expression of Kir4.1 in HEK293 cells transfected with Ki4.1+Nedd4-2, Kir4.1/5.1+Nedd4-2, Kir4.1+Nedd4-1, and Kir4.1/5.1+Nedd4-1. The expression of Nedd4 was shown in the middle panel. B: Normalized band density of Kir4.1 expression is shown in a bar graph. C: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents were measured (at −60 mV) with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1+Nedd4-2, Kir4.1/5.1T249A+Nedd4-2, and Kir4.1/5.1T249D +Nedd4-2, respectively (n = 6). D: Ba2+-sensitive K currents measured at −60 mV with whole-cell recording in HEK293 cells transfected with Kir4.1/5.1, Kir4.1/5.1T249A, and Kir4.1/5.1T249D (n = 6). Patch-clamp experiments were performed 24 h after the transfection, and symmetrical 145 mM K was used for the pipette and the bath solution. pA, picoamperes; pF, picofarads.

    Techniques Used: Mutagenesis, Inhibition, Western Blot, Expressing, Transfection, Patch Clamp, Transferring

    Deletion of Kir5.1 stimulates the basolateral Kir4.1 in the distal convoluted tubule (DCT). A: Whole-cell recording showing Ba2+-sensitive K currents in the DCT of the wild-type (WT) or Kir5.1 knockout (KO) mice. K currents were measured with a ramp protocol from −100 to 100 mV using symmetrical 145 mM K solution in the bath and pipette. B: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the DCT were measured at −60 mV from the WT and Kir5.1 KO mice (n = 6). Western blot (IB) shows the expression of Kir4.1 in WT and Kir5.1 KO mice (C), and a bar graph summarizes normalized band density of Kir4.1 (D). E: Immunostaining of Kir4.1 in kidney from WT and Kir5.1 KO mice. Treatment for the kidney slides was identical. pA, picoamperes.
    Figure Legend Snippet: Deletion of Kir5.1 stimulates the basolateral Kir4.1 in the distal convoluted tubule (DCT). A: Whole-cell recording showing Ba2+-sensitive K currents in the DCT of the wild-type (WT) or Kir5.1 knockout (KO) mice. K currents were measured with a ramp protocol from −100 to 100 mV using symmetrical 145 mM K solution in the bath and pipette. B: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the DCT were measured at −60 mV from the WT and Kir5.1 KO mice (n = 6). Western blot (IB) shows the expression of Kir4.1 in WT and Kir5.1 KO mice (C), and a bar graph summarizes normalized band density of Kir4.1 (D). E: Immunostaining of Kir4.1 in kidney from WT and Kir5.1 KO mice. Treatment for the kidney slides was identical. pA, picoamperes.

    Techniques Used: Knock-Out, Transferring, Western Blot, Expressing, Immunostaining

    Deletion of Nedd4-2 increases Kir4.1 expression. A: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the distal convoluted tubule (DCT) were measured at −60 mV from the wild-type (WT) and Ks-Nedd4-2 knockout (KO) mice (n = 6). B: Western blots (IB) showed the expression of Kir4.1 and Kir5.1 in WT and Ks-Nedd4-2 KO mice. A bar graph summarizes normalized band density of Kir4.1 and Ki5.1 (right panel). C: immunostaining of Kir4.1 in kidney from Ks-Nedd4-2 KO mice and WT control (without doxycycline treated). pA, picoamperes.
    Figure Legend Snippet: Deletion of Nedd4-2 increases Kir4.1 expression. A: Bar graph summarizes the results of experiments in which Ba2+-sensitive K currents of the distal convoluted tubule (DCT) were measured at −60 mV from the wild-type (WT) and Ks-Nedd4-2 knockout (KO) mice (n = 6). B: Western blots (IB) showed the expression of Kir4.1 and Kir5.1 in WT and Ks-Nedd4-2 KO mice. A bar graph summarizes normalized band density of Kir4.1 and Ki5.1 (right panel). C: immunostaining of Kir4.1 in kidney from Ks-Nedd4-2 KO mice and WT control (without doxycycline treated). pA, picoamperes.

    Techniques Used: Expressing, Knock-Out, Western Blot, Immunostaining

    A scheme illustrating the role of Kir5.1 as a binding partner in mediating Nedd4-2 E3 ligase dependent degradation of Kir4.1 in the distal convoluted tubule (DCT). A part of Kir5.1 sequence (from AA241 to 260) including TPVT motif is shown on the top. Ub, ubiquitin.
    Figure Legend Snippet: A scheme illustrating the role of Kir5.1 as a binding partner in mediating Nedd4-2 E3 ligase dependent degradation of Kir4.1 in the distal convoluted tubule (DCT). A part of Kir5.1 sequence (from AA241 to 260) including TPVT motif is shown on the top. Ub, ubiquitin.

    Techniques Used: Binding Assay, Sequencing

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    Alomone Labs kir4 1
    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and <t>Kir4.1</t> + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.
    Kir4 1, 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 kcnj10 antibodies
    AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). <t>Anti-KCNJ10</t> antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.
    Kcnj10 Antibodies, 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 apc 165
    AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). <t>Anti-KCNJ10</t> antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.
    Apc 165, 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 kir 4 1

    Kir 4 1, 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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    Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.

    Journal: Molecular Brain

    Article Title: Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes

    doi: 10.1186/1756-6606-6-28

    Figure Lengend Snippet: Impaired astrocytes were subsequently recovered in the LPS-injected SNpc. ( A - D ) LPS (5 μg/2 μl) was injected into the SNpc. Sections obtained at the indicated times after LPS injection were labeled with antibodies specific for GFAP ( A ), S100β ( B ), EAAT1 ( C ), and Kir 4.1 ( D ), and visualized with peroxidase-conjugated secondary antibodies. Lower panels (SNpc and SNr) are higher magnification images of boxed areas (SNpc and SNr) in upper panels. Arrows indicate GFAP + ( A ), S100β + ( B ) EAAT1 + ( C ) and Kir4.1 + ( D ) cells. Contralateral (Contra) sides of LPS-injected animals and PBS-injected animals were used as controls. Asterisks (*) indicate injection sites. Scale bars, 1 mm (upper panels in A ), 200 μm (upper panels in C - E ), 50 μm (lower panels in A ), 20 μm (lower panels in C - E ). ( E - H ) GFAP-, S100β-, EAAT1-, and Kir 4.1-negative areas in every sixth brain section were quantified using Axiovision image analysis software, as described in “Materials and Methods”. Values are means ± SEMs of at least three samples (*p < 0.01; **p < 0.001; Student’s t -test). Three or more animals were used for each time point. All data presented in this study are representative of at least three independent experiments unless indicated.

    Article Snippet: Kir4.1 , rabbit , 1:800 , apc-035 , Alomone.

    Techniques: Injection, Labeling, Software

    Primary antibodies used in immunohistochemistry

    Journal: Molecular Brain

    Article Title: Repair of astrocytes, blood vessels, and myelin in the injured brain: possible roles of blood monocytes

    doi: 10.1186/1756-6606-6-28

    Figure Lengend Snippet: Primary antibodies used in immunohistochemistry

    Article Snippet: Kir4.1 , rabbit , 1:800 , apc-035 , Alomone.

    Techniques:

    AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). Anti-KCNJ10 antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: AAV8BP2 and AAV8 transduce the mammalian cochlear lateral wall and endolymphatic sac with high efficiency

    doi: 10.1016/j.omtm.2022.07.013

    Figure Lengend Snippet: AAV8BP2-GFP and AAV8-GFP transduced the intermediate cells in the stria vascularis (A) Confocal images of the cochlear lateral wall at the level of the intermediate cell layer are shown (single optical section). Anti-KCNJ10 antibody was used as an intermediate cell marker. AAV8BP2-GFP (n = 9) and AAV8-GFP (n = 9) transduced the intermediate cells in the stria vascularis, but AAV2.7m8-GFP (n = 4), Anc80L65-GFP (n = 5), and AAV2-GFP (n = 4) showed minimal intermediate cell transduction. All images were taken at ∼P30. Surgeries were performed in P0–5 animals and the average surgery age was P1.9. The scale bar represents 50 μm for the lower-magnification images, and 10 μm for the magnified images. White dashed squares indicate areas where magnified images are taken. (B) Quantification of the intermediate cell transduction efficiency with various AAV serotypes. Both individual (open circles) and average results are presented. ∗p < 0.05. Error bars represent SEM.

    Article Snippet: For the primary antibodies, SLC12A2 antibodies were used to label marginal cells (1:100, Cat# sc21545; Santa Cruz Biotech, Dallas, TX), and KCNJ10 antibodies for intermediate cells (1:100, Cat# APC-035; Alomone Labs, Jerusalem, Israel).

    Techniques: Marker, Transduction

    Journal: eLife

    Article Title: Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit

    doi: 10.7554/eLife.71379

    Figure Lengend Snippet:

    Article Snippet: Antibody , Kir 4.1 extracellular (rabbit polyclonal) , Alomone Labs , APC-165 , Immunofluorescence (1:200).

    Techniques: Sequencing, Western Blot, Immunofluorescence, Transduction