anti aquaporin 5 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti aquaporin 5 antibody
    LAMP3 increases degradation of NKCC1 and <t>AQP5</t> by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P
    Anti Aquaporin 5 Antibody, supplied by Cell Signaling Technology Inc, 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/anti aquaporin 5 antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti aquaporin 5 antibody - by Bioz Stars, 2022-12
    94/100 stars

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    1) Product Images from "Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy"

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    Journal: Scientific Reports

    doi: 10.1038/s41598-022-21374-2

    LAMP3 increases degradation of NKCC1 and AQP5 by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P
    Figure Legend Snippet: LAMP3 increases degradation of NKCC1 and AQP5 by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P

    Techniques Used: In Vitro, Transfection, Expressing, Plasmid Preparation, Staining, Western Blot

    LAMP3 promotes degradation of NKCC1 and AQP5 in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells). ( A ) Representative cropped Western blots showing expression of NKCC1, AQP5, or α-tubulin (internal control) 48 h after transfection. ( B ) Relative change in SLC12A2 and AQP5 mRNA expressions to ACTB expression determined by real-time quantitative PCR 48 h after transfection. ( C ) Representative cropped Western blots after cells were treated with 50 μg/mL cycloheximide (CHX) for indicated hours. Graph shows remaining protein expression after 8 h relative to baseline. Values shown are mean ± SEM (n = 3 for all experiments). * P
    Figure Legend Snippet: LAMP3 promotes degradation of NKCC1 and AQP5 in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells). ( A ) Representative cropped Western blots showing expression of NKCC1, AQP5, or α-tubulin (internal control) 48 h after transfection. ( B ) Relative change in SLC12A2 and AQP5 mRNA expressions to ACTB expression determined by real-time quantitative PCR 48 h after transfection. ( C ) Representative cropped Western blots after cells were treated with 50 μg/mL cycloheximide (CHX) for indicated hours. Graph shows remaining protein expression after 8 h relative to baseline. Values shown are mean ± SEM (n = 3 for all experiments). * P

    Techniques Used: In Vitro, Transfection, Expressing, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction

    AAV2-AQP1 gene therapy restores salivary flow rate in LAMP3-overexpressing mice. ( A ) LAMP3 overexpression in submandibular glands of C57BL/6 mice was achieved by retrograde cannulation of AAV2-LAMP3. After 7 months, AAV2-AQP1 or AAV2-GFP (control) was delivered to submandibular glands of LAMP3-overexpressing mice by retrograde cannulation. ( B ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min in mice. ( C ) Representative immunofluorescent images of submandibular glands for NKCC1 (green), AQP5 (green), or AQP1 (magenta) (scale bars = 10 µm). ( D ) NKCC1 and AQP5 expression area in glands. ( E ) Serum anti-SSA/Ro antibody levels. ( F ) Size of lymphocyte infiltration area in glands. Values shown are mean ± SD (3 baseline, 5 AAV2-GFP-treated, and 4 AAV2-AQP1-treated mice). ** P
    Figure Legend Snippet: AAV2-AQP1 gene therapy restores salivary flow rate in LAMP3-overexpressing mice. ( A ) LAMP3 overexpression in submandibular glands of C57BL/6 mice was achieved by retrograde cannulation of AAV2-LAMP3. After 7 months, AAV2-AQP1 or AAV2-GFP (control) was delivered to submandibular glands of LAMP3-overexpressing mice by retrograde cannulation. ( B ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min in mice. ( C ) Representative immunofluorescent images of submandibular glands for NKCC1 (green), AQP5 (green), or AQP1 (magenta) (scale bars = 10 µm). ( D ) NKCC1 and AQP5 expression area in glands. ( E ) Serum anti-SSA/Ro antibody levels. ( F ) Size of lymphocyte infiltration area in glands. Values shown are mean ± SD (3 baseline, 5 AAV2-GFP-treated, and 4 AAV2-AQP1-treated mice). ** P

    Techniques Used: Mouse Assay, Over Expression, Expressing

    Graphical summary. LAMP3 overexpression decreases AQP5—a water channel in the apical membrane—and NKCC1—an ion cotransporter on the basolateral membrane—expressions in salivary gland epithelial cells by promoting endolysosomal degradation, resulting in salivary gland hypofunction. Simultaneously, LAMP3 increases internalization of AAV2 and enhances AAV2-mediated gene transduction efficiency via the activated endolysosomal pathway. AAV2-mediated AQP1 gene therapy induces ectopic AQP1 expression in both apical and basolateral membranes of salivary gland epithelial cells and creates a new pathway for fluid movement, leading to restoration of salivary flow rate in LAMP3-overexpressing mice.
    Figure Legend Snippet: Graphical summary. LAMP3 overexpression decreases AQP5—a water channel in the apical membrane—and NKCC1—an ion cotransporter on the basolateral membrane—expressions in salivary gland epithelial cells by promoting endolysosomal degradation, resulting in salivary gland hypofunction. Simultaneously, LAMP3 increases internalization of AAV2 and enhances AAV2-mediated gene transduction efficiency via the activated endolysosomal pathway. AAV2-mediated AQP1 gene therapy induces ectopic AQP1 expression in both apical and basolateral membranes of salivary gland epithelial cells and creates a new pathway for fluid movement, leading to restoration of salivary flow rate in LAMP3-overexpressing mice.

    Techniques Used: Over Expression, Transduction, Expressing, Mouse Assay

    LAMP3 causes salivary gland hypofunction through endolysosomal degradation of NKCC1 and AQP5 in vivo. Submandibular glands of C57BL/6 mice were instilled with AAV2-LAMP3 or AAV2-GFP (control), after which mice received weekly intraperitoneal injections of hydroxychloroquine (HCQ) at 60 mg/kg for 4 months. ( A ) Representative immunofluorescent images of submandibular glands showing staining for NKCC1 or AQP5 (both green) (scale bars = 20 µm). ( B , C ) Bar charts showing percentage of ( B ) NKCC1 and ( C ) AQP5 expressions per nuclear area in submandibular gland specimens. ( D ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min. Values shown are mean ± SD (4 control, 5 LAMP3-overexpressing, and 4 HCQ-treated LAMP3-overexpressing mice). ** P
    Figure Legend Snippet: LAMP3 causes salivary gland hypofunction through endolysosomal degradation of NKCC1 and AQP5 in vivo. Submandibular glands of C57BL/6 mice were instilled with AAV2-LAMP3 or AAV2-GFP (control), after which mice received weekly intraperitoneal injections of hydroxychloroquine (HCQ) at 60 mg/kg for 4 months. ( A ) Representative immunofluorescent images of submandibular glands showing staining for NKCC1 or AQP5 (both green) (scale bars = 20 µm). ( B , C ) Bar charts showing percentage of ( B ) NKCC1 and ( C ) AQP5 expressions per nuclear area in submandibular gland specimens. ( D ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min. Values shown are mean ± SD (4 control, 5 LAMP3-overexpressing, and 4 HCQ-treated LAMP3-overexpressing mice). ** P

    Techniques Used: In Vivo, Mouse Assay, Staining

    Decreased NKCC1 and AQP5 expressions are associated with salivary gland hypofunction in SjD patients. ( A ) Individuals who visited a rheumatology clinic because of sicca symptoms underwent labial minor salivary glands (MSGs) biopsy, measurement of salivary flow rate (SFR) and serum anti-Ro/SSA antibody tests. Thereafter, participants were divided into two groups: SjD patients with decreased SFR ( n = 5) and control subjects with normal SFR ( n = 6). ( B ) Representative immunofluorescent images of MSGs indicating staining for NKCC1 or AQP5 (green) and dot plots showing mean fluorescent intensity (MFI) of NKCC1 or AQP5 staining in each MSG (scale bars = 50 µm). Values shown are mean ± SD. * P
    Figure Legend Snippet: Decreased NKCC1 and AQP5 expressions are associated with salivary gland hypofunction in SjD patients. ( A ) Individuals who visited a rheumatology clinic because of sicca symptoms underwent labial minor salivary glands (MSGs) biopsy, measurement of salivary flow rate (SFR) and serum anti-Ro/SSA antibody tests. Thereafter, participants were divided into two groups: SjD patients with decreased SFR ( n = 5) and control subjects with normal SFR ( n = 6). ( B ) Representative immunofluorescent images of MSGs indicating staining for NKCC1 or AQP5 (green) and dot plots showing mean fluorescent intensity (MFI) of NKCC1 or AQP5 staining in each MSG (scale bars = 50 µm). Values shown are mean ± SD. * P

    Techniques Used: Staining

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    Cell Signaling Technology Inc anti aquaporin 5 antibody
    LAMP3 increases degradation of NKCC1 and <t>AQP5</t> by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P
    Anti Aquaporin 5 Antibody, supplied by Cell Signaling Technology Inc, 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/anti aquaporin 5 antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti aquaporin 5 antibody - by Bioz Stars, 2022-12
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    LAMP3 increases degradation of NKCC1 and AQP5 by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: LAMP3 increases degradation of NKCC1 and AQP5 by promoting endolysosomal degradation in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells) and 48 h thereafter treated with chloroquine (CQ) at 25 μM for 6 h. ( A ) Representative immunofluorescent images for LAMP3 (gray) or NKCC1 (red) and EEA1 (green) (scale bars = 5 µm). ( B – D ) Bar charts showing ( B ) relative change in EEA1 puncta area size, ( C ) Pearson correlation coefficients between EEA1 and NKCC1, and ( D ) relative change in mean fluorescent intensity (MFI) of NKCC1 staining ( n = 3 for all experiments). ( E ) Representative cropped Western blots showing expression of NKCC1, AQP5, and α-tubulin (internal control). Graph shows NKCC1 and AQP5 expression relative to empty ( n = 3). Values shown are mean ± SEM. ** P

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: In Vitro, Transfection, Expressing, Plasmid Preparation, Staining, Western Blot

    LAMP3 promotes degradation of NKCC1 and AQP5 in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells). ( A ) Representative cropped Western blots showing expression of NKCC1, AQP5, or α-tubulin (internal control) 48 h after transfection. ( B ) Relative change in SLC12A2 and AQP5 mRNA expressions to ACTB expression determined by real-time quantitative PCR 48 h after transfection. ( C ) Representative cropped Western blots after cells were treated with 50 μg/mL cycloheximide (CHX) for indicated hours. Graph shows remaining protein expression after 8 h relative to baseline. Values shown are mean ± SEM (n = 3 for all experiments). * P

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: LAMP3 promotes degradation of NKCC1 and AQP5 in vitro. HSG cells were transfected with LAMP3 expression or empty plasmid (3.0 μg per 1 × 10 6 cells). ( A ) Representative cropped Western blots showing expression of NKCC1, AQP5, or α-tubulin (internal control) 48 h after transfection. ( B ) Relative change in SLC12A2 and AQP5 mRNA expressions to ACTB expression determined by real-time quantitative PCR 48 h after transfection. ( C ) Representative cropped Western blots after cells were treated with 50 μg/mL cycloheximide (CHX) for indicated hours. Graph shows remaining protein expression after 8 h relative to baseline. Values shown are mean ± SEM (n = 3 for all experiments). * P

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: In Vitro, Transfection, Expressing, Plasmid Preparation, Western Blot, Real-time Polymerase Chain Reaction

    AAV2-AQP1 gene therapy restores salivary flow rate in LAMP3-overexpressing mice. ( A ) LAMP3 overexpression in submandibular glands of C57BL/6 mice was achieved by retrograde cannulation of AAV2-LAMP3. After 7 months, AAV2-AQP1 or AAV2-GFP (control) was delivered to submandibular glands of LAMP3-overexpressing mice by retrograde cannulation. ( B ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min in mice. ( C ) Representative immunofluorescent images of submandibular glands for NKCC1 (green), AQP5 (green), or AQP1 (magenta) (scale bars = 10 µm). ( D ) NKCC1 and AQP5 expression area in glands. ( E ) Serum anti-SSA/Ro antibody levels. ( F ) Size of lymphocyte infiltration area in glands. Values shown are mean ± SD (3 baseline, 5 AAV2-GFP-treated, and 4 AAV2-AQP1-treated mice). ** P

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: AAV2-AQP1 gene therapy restores salivary flow rate in LAMP3-overexpressing mice. ( A ) LAMP3 overexpression in submandibular glands of C57BL/6 mice was achieved by retrograde cannulation of AAV2-LAMP3. After 7 months, AAV2-AQP1 or AAV2-GFP (control) was delivered to submandibular glands of LAMP3-overexpressing mice by retrograde cannulation. ( B ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min in mice. ( C ) Representative immunofluorescent images of submandibular glands for NKCC1 (green), AQP5 (green), or AQP1 (magenta) (scale bars = 10 µm). ( D ) NKCC1 and AQP5 expression area in glands. ( E ) Serum anti-SSA/Ro antibody levels. ( F ) Size of lymphocyte infiltration area in glands. Values shown are mean ± SD (3 baseline, 5 AAV2-GFP-treated, and 4 AAV2-AQP1-treated mice). ** P

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: Mouse Assay, Over Expression, Expressing

    Graphical summary. LAMP3 overexpression decreases AQP5—a water channel in the apical membrane—and NKCC1—an ion cotransporter on the basolateral membrane—expressions in salivary gland epithelial cells by promoting endolysosomal degradation, resulting in salivary gland hypofunction. Simultaneously, LAMP3 increases internalization of AAV2 and enhances AAV2-mediated gene transduction efficiency via the activated endolysosomal pathway. AAV2-mediated AQP1 gene therapy induces ectopic AQP1 expression in both apical and basolateral membranes of salivary gland epithelial cells and creates a new pathway for fluid movement, leading to restoration of salivary flow rate in LAMP3-overexpressing mice.

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: Graphical summary. LAMP3 overexpression decreases AQP5—a water channel in the apical membrane—and NKCC1—an ion cotransporter on the basolateral membrane—expressions in salivary gland epithelial cells by promoting endolysosomal degradation, resulting in salivary gland hypofunction. Simultaneously, LAMP3 increases internalization of AAV2 and enhances AAV2-mediated gene transduction efficiency via the activated endolysosomal pathway. AAV2-mediated AQP1 gene therapy induces ectopic AQP1 expression in both apical and basolateral membranes of salivary gland epithelial cells and creates a new pathway for fluid movement, leading to restoration of salivary flow rate in LAMP3-overexpressing mice.

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: Over Expression, Transduction, Expressing, Mouse Assay

    LAMP3 causes salivary gland hypofunction through endolysosomal degradation of NKCC1 and AQP5 in vivo. Submandibular glands of C57BL/6 mice were instilled with AAV2-LAMP3 or AAV2-GFP (control), after which mice received weekly intraperitoneal injections of hydroxychloroquine (HCQ) at 60 mg/kg for 4 months. ( A ) Representative immunofluorescent images of submandibular glands showing staining for NKCC1 or AQP5 (both green) (scale bars = 20 µm). ( B , C ) Bar charts showing percentage of ( B ) NKCC1 and ( C ) AQP5 expressions per nuclear area in submandibular gland specimens. ( D ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min. Values shown are mean ± SD (4 control, 5 LAMP3-overexpressing, and 4 HCQ-treated LAMP3-overexpressing mice). ** P

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: LAMP3 causes salivary gland hypofunction through endolysosomal degradation of NKCC1 and AQP5 in vivo. Submandibular glands of C57BL/6 mice were instilled with AAV2-LAMP3 or AAV2-GFP (control), after which mice received weekly intraperitoneal injections of hydroxychloroquine (HCQ) at 60 mg/kg for 4 months. ( A ) Representative immunofluorescent images of submandibular glands showing staining for NKCC1 or AQP5 (both green) (scale bars = 20 µm). ( B , C ) Bar charts showing percentage of ( B ) NKCC1 and ( C ) AQP5 expressions per nuclear area in submandibular gland specimens. ( D ) Pilocarpine-stimulated salivary flow rate per body weight in 20 min. Values shown are mean ± SD (4 control, 5 LAMP3-overexpressing, and 4 HCQ-treated LAMP3-overexpressing mice). ** P

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: In Vivo, Mouse Assay, Staining

    Decreased NKCC1 and AQP5 expressions are associated with salivary gland hypofunction in SjD patients. ( A ) Individuals who visited a rheumatology clinic because of sicca symptoms underwent labial minor salivary glands (MSGs) biopsy, measurement of salivary flow rate (SFR) and serum anti-Ro/SSA antibody tests. Thereafter, participants were divided into two groups: SjD patients with decreased SFR ( n = 5) and control subjects with normal SFR ( n = 6). ( B ) Representative immunofluorescent images of MSGs indicating staining for NKCC1 or AQP5 (green) and dot plots showing mean fluorescent intensity (MFI) of NKCC1 or AQP5 staining in each MSG (scale bars = 50 µm). Values shown are mean ± SD. * P

    Journal: Scientific Reports

    Article Title: Correction of LAMP3-associated salivary gland hypofunction by aquaporin gene therapy

    doi: 10.1038/s41598-022-21374-2

    Figure Lengend Snippet: Decreased NKCC1 and AQP5 expressions are associated with salivary gland hypofunction in SjD patients. ( A ) Individuals who visited a rheumatology clinic because of sicca symptoms underwent labial minor salivary glands (MSGs) biopsy, measurement of salivary flow rate (SFR) and serum anti-Ro/SSA antibody tests. Thereafter, participants were divided into two groups: SjD patients with decreased SFR ( n = 5) and control subjects with normal SFR ( n = 6). ( B ) Representative immunofluorescent images of MSGs indicating staining for NKCC1 or AQP5 (green) and dot plots showing mean fluorescent intensity (MFI) of NKCC1 or AQP5 staining in each MSG (scale bars = 50 µm). Values shown are mean ± SD. * P

    Article Snippet: Membranes were blocked with 2% non-fat dried milk at 25 °C for 1 h and then incubated at 4 °C overnight with one of the following primary antibodies: anti-NKCC1 (#4828, Cell Signaling Technology), anti-AQP5 (#AQP-005, Alomone labs), anti-caveolin-1 (#3267, Cell Signaling Technology), anti-clathrin heavy chain (#4796, Cell Signaling Technology), anti-KIAA0319L/AAVR (#21,016-1-AP, Proteintech), or anti-α-tubulin (#T6199, Sigma-Aldrich).

    Techniques: Staining