anti aqp5  (Alomone Labs)


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    Alomone Labs anti aqp5
    Expression of known AT1 cell markers in rat microarray data. Log2 expression data were generated from microarray experiments: AT1-like cells differentiated in culture ( blue ) (Day 2-6), freshly isolated AT1 cells ( purple ), freshly isolated AT2 cells ( red ), and other tissues ( black ). Table at bottom right includes previously described AT1 cell–specific genes, their associated Illumina Probe IDs, and their FDR-corrected P values in this study. FDR adjustment is based on the number of tests shown for known genes. AGER, advanced glycosylation end product–specific receptor; <t>AQP5,</t> aquaporin 5; CAV, caveolin; FDR, false-discovery rate; ILMN, Illumina probe number; PDPN, podoplanin. **Indicates significantly greater in rat AT1 and AT1-like cells compared to all others.
    Anti Aqp5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    anti aqp5 - by Bioz Stars, 2022-07
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    Images

    1) Product Images from "Cross-Species Transcriptome Profiling Identifies New Alveolar Epithelial Type I Cell–Specific Genes"

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

    Journal: American Journal of Respiratory Cell and Molecular Biology

    doi: 10.1165/rcmb.2016-0071OC

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

    Techniques Used: Expressing, Microarray, Generated, Isolation

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

    Techniques Used: Expressing, Double Staining

    2) Product Images from "Localization of AQP5 during development of the mouse submandibular salivary gland"

    Article Title: Localization of AQP5 during development of the mouse submandibular salivary gland

    Journal: Journal of Molecular Histology

    doi: 10.1007/s10735-010-9308-0

    AQP5 expression pattern during postnatal development of the mouse submandibular gland (SMG). A At birth (postnatal day 0, P0), the pro-acinar cells ( arrow ) and the intercalated ducts ( ID ) are AQP5 positive. No AQP5 is detected in the intralobular duct ( IAD ) or interlobular duct ( IED ). B Pre-weaning (P5), both pro-acini ( arrow ) and intercalated ducts ( ID ) are AQP5 positive. No AQP5 is detected in the striated duct ( SD ). C Young adult females (P25): acini ( arrow ) and the proximal part of the intercalated duct ( ID ) are positive, while the granulated convoluted tubule ( GCT ) is negative. D Adult females (P60) show the same AQP5 pattern as in C . In addition, no AQP5 is detected in the transition from GCT to striated duct ( arrowhead ). E Adult males (P60): acini ( arrow ) and entire intercalated ducts ( ID ) are positive while the granulated convoluted tubule ( GCT ) is negative. F IgG negative control in young adult female (P25) tissue shows no unspecific staining in the acini ( arrow ), intercalated duct ( ID ), or in the granulated convoluted tubule ( GCT ). Granules in the granulated convoluted tubule are not seen using this method. A – F Scale bar 50 μm
    Figure Legend Snippet: AQP5 expression pattern during postnatal development of the mouse submandibular gland (SMG). A At birth (postnatal day 0, P0), the pro-acinar cells ( arrow ) and the intercalated ducts ( ID ) are AQP5 positive. No AQP5 is detected in the intralobular duct ( IAD ) or interlobular duct ( IED ). B Pre-weaning (P5), both pro-acini ( arrow ) and intercalated ducts ( ID ) are AQP5 positive. No AQP5 is detected in the striated duct ( SD ). C Young adult females (P25): acini ( arrow ) and the proximal part of the intercalated duct ( ID ) are positive, while the granulated convoluted tubule ( GCT ) is negative. D Adult females (P60) show the same AQP5 pattern as in C . In addition, no AQP5 is detected in the transition from GCT to striated duct ( arrowhead ). E Adult males (P60): acini ( arrow ) and entire intercalated ducts ( ID ) are positive while the granulated convoluted tubule ( GCT ) is negative. F IgG negative control in young adult female (P25) tissue shows no unspecific staining in the acini ( arrow ), intercalated duct ( ID ), or in the granulated convoluted tubule ( GCT ). Granules in the granulated convoluted tubule are not seen using this method. A – F Scale bar 50 μm

    Techniques Used: Expressing, Negative Control, Staining

    Gold labeling of AQP5 in the SMG of adult animals (P60). A Overview of AQP5 gold staining in the basal membrane, membranes of intercellular canaliculi, as well as the lateral membrane ( LM ). CL canalicular lumen, BD basal digits (scale bar, 1 μm). B Gold labeling of AQP5 was detected in the apical membrane of acinar cells (scale bar, 1 μm). C Longitudinal section of an intercellular canaliculus showing gold labeling of AQP5 in the membrane (scale bar, 0.5 μm). D AQP5 was localized in the basal membrane in areas where digits could be seen, while no AQP5 was detectable in the basement membrane (scale bar 0.2 μm). Non-linear adjustments were applied to entire images
    Figure Legend Snippet: Gold labeling of AQP5 in the SMG of adult animals (P60). A Overview of AQP5 gold staining in the basal membrane, membranes of intercellular canaliculi, as well as the lateral membrane ( LM ). CL canalicular lumen, BD basal digits (scale bar, 1 μm). B Gold labeling of AQP5 was detected in the apical membrane of acinar cells (scale bar, 1 μm). C Longitudinal section of an intercellular canaliculus showing gold labeling of AQP5 in the membrane (scale bar, 0.5 μm). D AQP5 was localized in the basal membrane in areas where digits could be seen, while no AQP5 was detectable in the basement membrane (scale bar 0.2 μm). Non-linear adjustments were applied to entire images

    Techniques Used: Labeling, Staining

    Confocal imaging of the adult SMG (P60). Aquaporin 5 (AQP5) demonstrates an apical, lateral, and basal localization in the SMG acini. A Localization of ZO-1 ( green ) near luminal membranes of acinar cells. B Localization of AQP5 ( red ) within the same cells as in A . C Merged image of A and B confirms the apical localization of AQP5. Arrow heads luminal membrane (scale bar 20 μm). D Localization of E-cadherin ( green ) within lateral membranes in both acinar ( arrow heads ) and GCT cells (*). E Localization of AQP5 ( red ) in the acinar cells. F Merged image of D and E demonstrates a lateral localization of AQP5 ( yellow / orange ), designated by arrowheads . An AQP5 and E-cadherin non-overlapping pattern of expression in the luminal canaliculi is evident ( arrow ). Non-overlapping patterns are also seen in the GCT cells (*) (scale bar, 50 μm). G Immunostaining of the basement membrane protein coll IV (blue). H Localization of AQP5 ( red ) in the acinar cells. I Merged image confirms a basal localization ( arrow head ) of AQP5 distinct from coll IV localization (scale bar 20 μm). J Localization of E-cadherin ( green ) within lateral membranes in acinar cells in a tissue immunostained with preabsorbed AQP5. K Preabsorption of the AQP5 antibody using its cognate peptide, showing complete elimination of the AQP5 staining pattern in the same cells as in J (scale bar 20 μm). A – I images are of male P60, J – K images are of female P60
    Figure Legend Snippet: Confocal imaging of the adult SMG (P60). Aquaporin 5 (AQP5) demonstrates an apical, lateral, and basal localization in the SMG acini. A Localization of ZO-1 ( green ) near luminal membranes of acinar cells. B Localization of AQP5 ( red ) within the same cells as in A . C Merged image of A and B confirms the apical localization of AQP5. Arrow heads luminal membrane (scale bar 20 μm). D Localization of E-cadherin ( green ) within lateral membranes in both acinar ( arrow heads ) and GCT cells (*). E Localization of AQP5 ( red ) in the acinar cells. F Merged image of D and E demonstrates a lateral localization of AQP5 ( yellow / orange ), designated by arrowheads . An AQP5 and E-cadherin non-overlapping pattern of expression in the luminal canaliculi is evident ( arrow ). Non-overlapping patterns are also seen in the GCT cells (*) (scale bar, 50 μm). G Immunostaining of the basement membrane protein coll IV (blue). H Localization of AQP5 ( red ) in the acinar cells. I Merged image confirms a basal localization ( arrow head ) of AQP5 distinct from coll IV localization (scale bar 20 μm). J Localization of E-cadherin ( green ) within lateral membranes in acinar cells in a tissue immunostained with preabsorbed AQP5. K Preabsorption of the AQP5 antibody using its cognate peptide, showing complete elimination of the AQP5 staining pattern in the same cells as in J (scale bar 20 μm). A – I images are of male P60, J – K images are of female P60

    Techniques Used: Imaging, Expressing, Immunostaining, Staining

    AQP5 expression pattern during prenatal development of the mouse submandibular gland (SMG). A Pseudoglandular stage (~E14): no AQP5 staining is detectable in the terminal bud ( t ), epithelial stalk ( e ) or mesechyme ( m ). B Early canalicular stage (~E14-15): AQP5 negative cells in the terminal bud ( t ), presumptive duct ( pd ), and mesenchyme ( m ). C Late canalicular stage (~E15-E16): scattered positive pro-acinar cells are present ( arrow ). No AQP5 is found in the presumptive duct ( pd ) or mesenchyme ( m ). D Early terminal bud stage (~E16-E17): all pro-acinar cells are AQP5 positive ( arrow ). In the intralobular duct ( IAD ), cells proximal to the pro-acini show apical AQP5 staining ( arrowhead ). E and F Late terminal bud stage (~E17-18): a similar expression pattern is observed as in D . Additionally, the rest of the intralobular duct (IAD) is also positive, and the interlobular duct ( IED ) is negative. Scale bar ( A – E ) 50 μm and ( F ) 100 μm
    Figure Legend Snippet: AQP5 expression pattern during prenatal development of the mouse submandibular gland (SMG). A Pseudoglandular stage (~E14): no AQP5 staining is detectable in the terminal bud ( t ), epithelial stalk ( e ) or mesechyme ( m ). B Early canalicular stage (~E14-15): AQP5 negative cells in the terminal bud ( t ), presumptive duct ( pd ), and mesenchyme ( m ). C Late canalicular stage (~E15-E16): scattered positive pro-acinar cells are present ( arrow ). No AQP5 is found in the presumptive duct ( pd ) or mesenchyme ( m ). D Early terminal bud stage (~E16-E17): all pro-acinar cells are AQP5 positive ( arrow ). In the intralobular duct ( IAD ), cells proximal to the pro-acini show apical AQP5 staining ( arrowhead ). E and F Late terminal bud stage (~E17-18): a similar expression pattern is observed as in D . Additionally, the rest of the intralobular duct (IAD) is also positive, and the interlobular duct ( IED ) is negative. Scale bar ( A – E ) 50 μm and ( F ) 100 μm

    Techniques Used: Expressing, Staining

    3) Product Images from "Aquaporin gene therapy corrects Sjögren’s syndrome phenotype in mice"

    Article Title: Aquaporin gene therapy corrects Sjögren’s syndrome phenotype in mice

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1601992113

    BMP6-Induced Loss of Cellular Water Permeability Is Associated with AQP5 Down-Regulation.
    Figure Legend Snippet: BMP6-Induced Loss of Cellular Water Permeability Is Associated with AQP5 Down-Regulation.

    Techniques Used: Permeability

    BMP6 and AQP5 expression in MSGs from patients with pSS and HVs. Stitched immunofluorescence images of BMP6 and AQP5 in the entire biopsied sample area are shown. (Scale bar, 300 μm.)
    Figure Legend Snippet: BMP6 and AQP5 expression in MSGs from patients with pSS and HVs. Stitched immunofluorescence images of BMP6 and AQP5 in the entire biopsied sample area are shown. (Scale bar, 300 μm.)

    Techniques Used: Expressing, Immunofluorescence

    BMP6 inhibits AQP5 expression in patients with Sjögren’s syndrome and HSG cells. ( A ) HSG cells were cultured without ( Upper ) or with ( Lower ) 6 ng/mL BMP6 for 3 d. Actin expression was detected with phalloidin conjugated to TRITC (red fluorescence).
    Figure Legend Snippet: BMP6 inhibits AQP5 expression in patients with Sjögren’s syndrome and HSG cells. ( A ) HSG cells were cultured without ( Upper ) or with ( Lower ) 6 ng/mL BMP6 for 3 d. Actin expression was detected with phalloidin conjugated to TRITC (red fluorescence).

    Techniques Used: Expressing, Cell Culture, Fluorescence

    Aquaporins restore water permeability in BMP6 treated cells. HSG cells were placed in the hypotonic solution following culture without (purple line) or with 6 ng/mL (red line) BMP6 and then transfected with increasing amounts of DNA encoding AQP5 ( A and
    Figure Legend Snippet: Aquaporins restore water permeability in BMP6 treated cells. HSG cells were placed in the hypotonic solution following culture without (purple line) or with 6 ng/mL (red line) BMP6 and then transfected with increasing amounts of DNA encoding AQP5 ( A and

    Techniques Used: Permeability, Transfection

    4) Product Images from "Identification of the protective mechanisms of Lactoferrin in the irradiated salivary gland"

    Article Title: Identification of the protective mechanisms of Lactoferrin in the irradiated salivary gland

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-10351-9

    Effect of LF on irradiated SMGs in ex vivo organ culture. Phase-contrast images show E12.5 + 24 h cultured SMGs at 0 and 48 h of culture with 0.1 mg/ml LF ( a – d ). Phase-contrast images show E12.5 + 24 h cultured SMGs at 0 and 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) ( e – h ). Scale Bar: 500 µm. The effects of treatment with 0.1 mg/ml LF were quantified by counting the number of buds per gland at 24, 48 h ( i ) and at 24, 48 h after irradiation ( j ). Hematoxylin-Eosin staining images show E12.5 + 24 h cultured SMGs at 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) ( k – r ). Scale bar: 50 µm. ( m and n ) are higher magnifications of ( k and l ), respectively. ( o and p ) are higher magnifications of ( m and n ), respectively. Black area of ( q and r ) is intercellular space of ( o and p ). Quantification of the percent area per field (50 µm 2 /field) occupied by intercellular space is graphed (s). The expression of AQP5 mRNA in E12.5 + 24 h cultured SMGs at 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) was determined by qPCR analysis (t). Bars represent the mean ± SEM. The study was repeated three times, with data shown from a representative experiment. ** p
    Figure Legend Snippet: Effect of LF on irradiated SMGs in ex vivo organ culture. Phase-contrast images show E12.5 + 24 h cultured SMGs at 0 and 48 h of culture with 0.1 mg/ml LF ( a – d ). Phase-contrast images show E12.5 + 24 h cultured SMGs at 0 and 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) ( e – h ). Scale Bar: 500 µm. The effects of treatment with 0.1 mg/ml LF were quantified by counting the number of buds per gland at 24, 48 h ( i ) and at 24, 48 h after irradiation ( j ). Hematoxylin-Eosin staining images show E12.5 + 24 h cultured SMGs at 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) ( k – r ). Scale bar: 50 µm. ( m and n ) are higher magnifications of ( k and l ), respectively. ( o and p ) are higher magnifications of ( m and n ), respectively. Black area of ( q and r ) is intercellular space of ( o and p ). Quantification of the percent area per field (50 µm 2 /field) occupied by intercellular space is graphed (s). The expression of AQP5 mRNA in E12.5 + 24 h cultured SMGs at 48 h of culture with 0.1 mg/ml LF after irradiation (4 Gy) was determined by qPCR analysis (t). Bars represent the mean ± SEM. The study was repeated three times, with data shown from a representative experiment. ** p

    Techniques Used: Irradiation, Ex Vivo, Organ Culture, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction

    Effect of LF on irradiated salivary gland in vivo . Hematoxylin-Eosin staining images show salivary glands from 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 6) ( a – d ). Scale bar: 100 µm. ( c and d ) are higher magnifications of ( a and b ), respectively. ( f and g ) are higher magnifications of ( c and d ), respectively. Quantification of the percent areas per field (250 µm 2 /field) occupied by intercellular space and by acinar cells is graphed ( e and h ) ( n = 3). The expression of AQP5 mRNA in 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) was determined by qPCR analysis ( n = 9) ( i ). Immunofluorescent images show the localization of AQP5, E-Cadherin, and DAPI in salivary glands from 6-week old mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 4) ( j , k ). Scale bar: 50 µm. The total pixel intensity of AQP5 at 48 h was quantified using NIH ImageJ software and was normalized to E-Cadherin ( l ). Salivary amylase activity was examined by the Assay Kit from 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 3) ( m ). Water intake was examined using water supply bottles from 6-week mice treated with 4.0 mg/animal LF at 1 day after irradiation (9 Gy) ( n = 12) ( n ). Bars represent the mean ± SEM. ** p
    Figure Legend Snippet: Effect of LF on irradiated salivary gland in vivo . Hematoxylin-Eosin staining images show salivary glands from 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 6) ( a – d ). Scale bar: 100 µm. ( c and d ) are higher magnifications of ( a and b ), respectively. ( f and g ) are higher magnifications of ( c and d ), respectively. Quantification of the percent areas per field (250 µm 2 /field) occupied by intercellular space and by acinar cells is graphed ( e and h ) ( n = 3). The expression of AQP5 mRNA in 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) was determined by qPCR analysis ( n = 9) ( i ). Immunofluorescent images show the localization of AQP5, E-Cadherin, and DAPI in salivary glands from 6-week old mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 4) ( j , k ). Scale bar: 50 µm. The total pixel intensity of AQP5 at 48 h was quantified using NIH ImageJ software and was normalized to E-Cadherin ( l ). Salivary amylase activity was examined by the Assay Kit from 6-week mice treated with 4.0 mg/animal LF at 1 week after irradiation (9 Gy) ( n = 3) ( m ). Water intake was examined using water supply bottles from 6-week mice treated with 4.0 mg/animal LF at 1 day after irradiation (9 Gy) ( n = 12) ( n ). Bars represent the mean ± SEM. ** p

    Techniques Used: Irradiation, In Vivo, Staining, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Software, Activity Assay

    5) Product Images from "Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage"

    Article Title: Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0141862

    In vitro effect of the hAdMSC secretome on cell death and proliferation of human parotid epithelial cells (HPEC) treated with the hAdMSC secretome at concentrations of 10, 50 and 100% or a normoxic control medium. (A–B) Light microscopic and immunofluorescent staining of HPEC. Scale bars represent 100 and 8μm in A and 100 and 20 μm in B. (C-D) Salivary epithelial genes of AQP5 and CK7 were confirmed by real-time PCR and western blot. *, compared to 2D, *P
    Figure Legend Snippet: In vitro effect of the hAdMSC secretome on cell death and proliferation of human parotid epithelial cells (HPEC) treated with the hAdMSC secretome at concentrations of 10, 50 and 100% or a normoxic control medium. (A–B) Light microscopic and immunofluorescent staining of HPEC. Scale bars represent 100 and 8μm in A and 100 and 20 μm in B. (C-D) Salivary epithelial genes of AQP5 and CK7 were confirmed by real-time PCR and western blot. *, compared to 2D, *P

    Techniques Used: In Vitro, Staining, Real-time Polymerase Chain Reaction, Western Blot

    6) Product Images from "Cross-Species Transcriptome Profiling Identifies New Alveolar Epithelial Type I Cell–Specific Genes"

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

    Journal: American Journal of Respiratory Cell and Molecular Biology

    doi: 10.1165/rcmb.2016-0071OC

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

    Techniques Used: Expressing, Microarray, Generated, Isolation

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

    Techniques Used: Expressing, Double Staining

    7) Product Images from "Integrated Transcriptomic and Epigenomic Analysis of Primary Human Lung Epithelial Cell Differentiation"

    Article Title: Integrated Transcriptomic and Epigenomic Analysis of Primary Human Lung Epithelial Cell Differentiation

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1003513

    Transcriptomic profiling of human AEC differentiation. A) Heatmap of top 5% variant-VSN normalized gene expression probes. Blue = low expression, red = high expression. DAY = number of days AT2 cells were allowed to differentiate. “Prep” = donor lung origin by color ( Figure S1 ). B) Principal component analysis of normalized hAEC samples. Samples color coded by donor lung as in (A). C) Significant changes in hAEC gene expression. Black line = BH-adjusted cutoff (FDR adjusted p≤0.05) calculated between D0 and D8. 20 genes show both significant up and downregulation for probes in different locations of the gene. D) Manhattan plot of differentially expressed genes. X-axis = chromosomal location, Y-axis = number of genes in each 2 MB region. E) qRT-PCR validation of microarray, data expressed in log 2 -fold change of differences between D0 and D8. Circles = top 10 up- and down-regulated genes, triangles = known AT1 cell differentiation markers ( AQP5, PDPN, CAV1 ). F) IPA of significantly up- or down-regulated genes. Bars expressed as log 10 -BH corrected p-values of enrichment for pathway members in significant list against RefSeq db38 background. Whole figure: Red = upregulated, green = downregulated.
    Figure Legend Snippet: Transcriptomic profiling of human AEC differentiation. A) Heatmap of top 5% variant-VSN normalized gene expression probes. Blue = low expression, red = high expression. DAY = number of days AT2 cells were allowed to differentiate. “Prep” = donor lung origin by color ( Figure S1 ). B) Principal component analysis of normalized hAEC samples. Samples color coded by donor lung as in (A). C) Significant changes in hAEC gene expression. Black line = BH-adjusted cutoff (FDR adjusted p≤0.05) calculated between D0 and D8. 20 genes show both significant up and downregulation for probes in different locations of the gene. D) Manhattan plot of differentially expressed genes. X-axis = chromosomal location, Y-axis = number of genes in each 2 MB region. E) qRT-PCR validation of microarray, data expressed in log 2 -fold change of differences between D0 and D8. Circles = top 10 up- and down-regulated genes, triangles = known AT1 cell differentiation markers ( AQP5, PDPN, CAV1 ). F) IPA of significantly up- or down-regulated genes. Bars expressed as log 10 -BH corrected p-values of enrichment for pathway members in significant list against RefSeq db38 background. Whole figure: Red = upregulated, green = downregulated.

    Techniques Used: Variant Assay, Expressing, Quantitative RT-PCR, Microarray, Cell Differentiation, Indirect Immunoperoxidase Assay

    Functional validation of a transcription factor signaling pathway predicted from bioinformatics analysis. A) Western blots examining AT2 and AT1 cell markers during differentiation in the presence or absence of RXR antagonist UVI-3003. LAMIN A/C is the loading control. B) Transepithelial resistance as measured in kΩ-cm 2 over the course of differentiation. Error bars represent technical duplicates for each plating. C) Rat Aqp5 -luciferase 4.3 kb promoter construct. Grey lines = 34 putative PPARA:RXR binding sites (Explain3.0). No sites were predicted from −900 to +6 bp due to lack of rat sequence information in the Explain v3.0 database. The asterisk marks the approximate location in the promoter of the ChIPed RXR site in E, below. The average number of PPARA:RXR sites per kilobase in the listed human/rat/mouse promoters is given in the table, with consensus site listed at the top. D) MLE-15 cells were transiently transfected with the Aqp5 -luciferase construct and treated for 48 hours with vehicle (DMSO) or 7.5 µM UVI-3003. UV1-3003 treatment reduced Aqp5 -luc activity by 48%±0.06. Values were normalized to vehicle control and represent the mean, error bars represent SEM, N = 3. All experiments represent 3 biological replicates. E) ChIP was performed on primary cultured rat AEC at day 0 (AT2, D0, n = 2) and day 8 (AT1-like, D8, n = 3). A region ∼4 kb upstream of the transcription start site specifically precipitated with RXR in day 8 samples. ChIP of GAPDH with RXR was performed as a control, and POL2 (POLR2A) binding to the GAPDH promoter was included as a positive control for the quality of day 0 DNA.
    Figure Legend Snippet: Functional validation of a transcription factor signaling pathway predicted from bioinformatics analysis. A) Western blots examining AT2 and AT1 cell markers during differentiation in the presence or absence of RXR antagonist UVI-3003. LAMIN A/C is the loading control. B) Transepithelial resistance as measured in kΩ-cm 2 over the course of differentiation. Error bars represent technical duplicates for each plating. C) Rat Aqp5 -luciferase 4.3 kb promoter construct. Grey lines = 34 putative PPARA:RXR binding sites (Explain3.0). No sites were predicted from −900 to +6 bp due to lack of rat sequence information in the Explain v3.0 database. The asterisk marks the approximate location in the promoter of the ChIPed RXR site in E, below. The average number of PPARA:RXR sites per kilobase in the listed human/rat/mouse promoters is given in the table, with consensus site listed at the top. D) MLE-15 cells were transiently transfected with the Aqp5 -luciferase construct and treated for 48 hours with vehicle (DMSO) or 7.5 µM UVI-3003. UV1-3003 treatment reduced Aqp5 -luc activity by 48%±0.06. Values were normalized to vehicle control and represent the mean, error bars represent SEM, N = 3. All experiments represent 3 biological replicates. E) ChIP was performed on primary cultured rat AEC at day 0 (AT2, D0, n = 2) and day 8 (AT1-like, D8, n = 3). A region ∼4 kb upstream of the transcription start site specifically precipitated with RXR in day 8 samples. ChIP of GAPDH with RXR was performed as a control, and POL2 (POLR2A) binding to the GAPDH promoter was included as a positive control for the quality of day 0 DNA.

    Techniques Used: Functional Assay, Western Blot, Luciferase, Construct, Binding Assay, Sequencing, Transfection, Activity Assay, Chromatin Immunoprecipitation, Cell Culture, Positive Control

    8) Product Images from "Fucoidan attenuates radioiodine-induced salivary gland dysfunction in mice"

    Article Title: Fucoidan attenuates radioiodine-induced salivary gland dysfunction in mice

    Journal: BMC Oral Health

    doi: 10.1186/s12903-019-0894-2

    Immunohistochemistry and TUNEL assay of salivary glands at 12 weeks post-RI treatment. Representative immunohistochemical images show salivary epithelial cells (aquaporins 5 (AQP5) staining) and myoepithelial cells (α-smooth muscle actin (α-SMA) staining). The expression level of AQP5 and α-SMA was lower in the radioiodine-exposed group (RI group) than in the normal group. Treatment with fucoidan increased the staining intensity of these cells relative to the RI group. The total number of TUNEL-positive cells was significantly reduced in the fucoidan group as compared with the RI group at 2, 4, and 12 weeks post RI. Statistical analyses were conducted using the Kruskal-Wallis test and the Dunn’s post hoc multiple comparison test. Each bar shows the mean ± SD; * , compared to the normal control group; # , compared to the RI group. ** p
    Figure Legend Snippet: Immunohistochemistry and TUNEL assay of salivary glands at 12 weeks post-RI treatment. Representative immunohistochemical images show salivary epithelial cells (aquaporins 5 (AQP5) staining) and myoepithelial cells (α-smooth muscle actin (α-SMA) staining). The expression level of AQP5 and α-SMA was lower in the radioiodine-exposed group (RI group) than in the normal group. Treatment with fucoidan increased the staining intensity of these cells relative to the RI group. The total number of TUNEL-positive cells was significantly reduced in the fucoidan group as compared with the RI group at 2, 4, and 12 weeks post RI. Statistical analyses were conducted using the Kruskal-Wallis test and the Dunn’s post hoc multiple comparison test. Each bar shows the mean ± SD; * , compared to the normal control group; # , compared to the RI group. ** p

    Techniques Used: Immunohistochemistry, TUNEL Assay, Staining, Expressing

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    Alomone Labs rabbit polyclonal anti aqp5 antibody
    Spatiotemporal expression of CXC-chemokine receptor 4 (CXCR4) and its ligand, CXCL12, and developmental genes in embryonic organs. ( A ) Schematic diagram of embryonic submandibular gland (eSMG) isolation and ex vivo culture. ( B ) Ex vivo branching morphogenesis of eSMGs from embryonic day (E) 13 to 17, showing epithelial growth and retraction of mesenchyme. Scale bars: 500 µm. ( C ) Temporal mRNA expression patterns of keratin 7 ( Krt7 ), aquaporin 5 ( <t>Aqp5</t> ), e-cadherin ( Cdh1 ), Krt15 , Cxcr4 , and Cxcl12 were measured from E13 to E17 by qPCR ( n = 3). ( D ) Epithelial (Epi) and mesenchymal (Mes) expression of Cxcr4 , Cxcl12 , odd-skipped related transcription factor 1 ( Osr1 ), and Cdh1 were quantified by qPCR at E13. The comparative C t values are expressed as fold increase relative to the epithelium ( n = 3). ( E ) Representative images showing expression of CXCR4 and CXCL12 in eSMG (upper) and their colocalization (lower) ( n = 3, scale bar: 500 µm). ( F ) Representative immunofluorescence images of CXCR4 and CXCL12 expression in E12 embryonic lung and pancreas ( n = 4); whole view (left two panels; scale bar: 500 µm) and magnified lumen structures (right two panels; scale bar: 50 µm). Data are presented as the mean ± SEM; * p
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    Spatiotemporal expression of CXC-chemokine receptor 4 (CXCR4) and its ligand, CXCL12, and developmental genes in embryonic organs. ( A ) Schematic diagram of embryonic submandibular gland (eSMG) isolation and ex vivo culture. ( B ) Ex vivo branching morphogenesis of eSMGs from embryonic day (E) 13 to 17, showing epithelial growth and retraction of mesenchyme. Scale bars: 500 µm. ( C ) Temporal mRNA expression patterns of keratin 7 ( Krt7 ), aquaporin 5 ( Aqp5 ), e-cadherin ( Cdh1 ), Krt15 , Cxcr4 , and Cxcl12 were measured from E13 to E17 by qPCR ( n = 3). ( D ) Epithelial (Epi) and mesenchymal (Mes) expression of Cxcr4 , Cxcl12 , odd-skipped related transcription factor 1 ( Osr1 ), and Cdh1 were quantified by qPCR at E13. The comparative C t values are expressed as fold increase relative to the epithelium ( n = 3). ( E ) Representative images showing expression of CXCR4 and CXCL12 in eSMG (upper) and their colocalization (lower) ( n = 3, scale bar: 500 µm). ( F ) Representative immunofluorescence images of CXCR4 and CXCL12 expression in E12 embryonic lung and pancreas ( n = 4); whole view (left two panels; scale bar: 500 µm) and magnified lumen structures (right two panels; scale bar: 50 µm). Data are presented as the mean ± SEM; * p

    Journal: International Journal of Molecular Sciences

    Article Title: CXCR4 Regulates Temporal Differentiation via PRC1 Complex in Organogenesis of Epithelial Glands

    doi: 10.3390/ijms22020619

    Figure Lengend Snippet: Spatiotemporal expression of CXC-chemokine receptor 4 (CXCR4) and its ligand, CXCL12, and developmental genes in embryonic organs. ( A ) Schematic diagram of embryonic submandibular gland (eSMG) isolation and ex vivo culture. ( B ) Ex vivo branching morphogenesis of eSMGs from embryonic day (E) 13 to 17, showing epithelial growth and retraction of mesenchyme. Scale bars: 500 µm. ( C ) Temporal mRNA expression patterns of keratin 7 ( Krt7 ), aquaporin 5 ( Aqp5 ), e-cadherin ( Cdh1 ), Krt15 , Cxcr4 , and Cxcl12 were measured from E13 to E17 by qPCR ( n = 3). ( D ) Epithelial (Epi) and mesenchymal (Mes) expression of Cxcr4 , Cxcl12 , odd-skipped related transcription factor 1 ( Osr1 ), and Cdh1 were quantified by qPCR at E13. The comparative C t values are expressed as fold increase relative to the epithelium ( n = 3). ( E ) Representative images showing expression of CXCR4 and CXCL12 in eSMG (upper) and their colocalization (lower) ( n = 3, scale bar: 500 µm). ( F ) Representative immunofluorescence images of CXCR4 and CXCL12 expression in E12 embryonic lung and pancreas ( n = 4); whole view (left two panels; scale bar: 500 µm) and magnified lumen structures (right two panels; scale bar: 50 µm). Data are presented as the mean ± SEM; * p

    Article Snippet: The following primary antibodies were used in the procedures: rat monoclonal anti-CXCR4 antibody (R & D Systems, MAB21651), rabbit monoclonal anti-KRT7 antibody (Abcam, ab181598; Cambridge, UK); rabbit monoclonal anti-CDH1 antibody (CST, 3195; Beverly, MA, USA), rabbit monoclonal anti-H2AK119ub antibody (CST, 8240); mouse monoclonal anti-CXCL12 antibody (Novus Biologicals, MAB350; Littleton, CO, USA), rabbit polyclonal anti-AQP5 antibody (Alomone Labs, AQP-005; Jerusalem, Israel), rabbit monoclonal anti-CASP3 antibody (CST, 9664), and rat monoclonal anti-KI67 antibody (ThermoFisher, 14-5698-82).

    Techniques: Expressing, Isolation, Ex Vivo, Real-time Polymerase Chain Reaction, Immunofluorescence

    AMD3100-induced precocious differentiation of epithelial cells. ( A , B ) Representative contour tracing ( A ) and bud number changes ( B ) of control and AMD3100-treated eSMGs during 48 h at 6-h intervals ( n = 3). ( C ) EdU staining results at 6 and 24 h after AMD3100 treatment. EdU in green and PNA in gray ( n = 4, scale bar: 500 µm). ( D ) Immunostaining results of Ki67 (red) and F-actin (green) in acini and duct of eSMGs 24 h after AMD3100 treatment. Morphologies of acinar buds and duct cells are outlined with white dotted lines. Scale bar: 50 µm. ( E ) Duct widths of control and AMD3100-treated eSMGs were visualized via F-actin-based intensity profiles of horizontal sectioning of ducts 24 h after the treatment. ( F ) Duct widths of control and AMD3100-treated eSMGs were quantified 24 h after the treatment ( n = 9). ( G ) Immunostaining results of AQP5 (green) and KRT7 (red). Magnified regions of acinar buds are marked with white dotted squares. The white arrows (middle panels) indicate areas with the highest AQP5 expression ( n = 4, scale bar: left, 100 µm; middle and right, 50 µm). Data are presented as the mean ± SEM; * p

    Journal: International Journal of Molecular Sciences

    Article Title: CXCR4 Regulates Temporal Differentiation via PRC1 Complex in Organogenesis of Epithelial Glands

    doi: 10.3390/ijms22020619

    Figure Lengend Snippet: AMD3100-induced precocious differentiation of epithelial cells. ( A , B ) Representative contour tracing ( A ) and bud number changes ( B ) of control and AMD3100-treated eSMGs during 48 h at 6-h intervals ( n = 3). ( C ) EdU staining results at 6 and 24 h after AMD3100 treatment. EdU in green and PNA in gray ( n = 4, scale bar: 500 µm). ( D ) Immunostaining results of Ki67 (red) and F-actin (green) in acini and duct of eSMGs 24 h after AMD3100 treatment. Morphologies of acinar buds and duct cells are outlined with white dotted lines. Scale bar: 50 µm. ( E ) Duct widths of control and AMD3100-treated eSMGs were visualized via F-actin-based intensity profiles of horizontal sectioning of ducts 24 h after the treatment. ( F ) Duct widths of control and AMD3100-treated eSMGs were quantified 24 h after the treatment ( n = 9). ( G ) Immunostaining results of AQP5 (green) and KRT7 (red). Magnified regions of acinar buds are marked with white dotted squares. The white arrows (middle panels) indicate areas with the highest AQP5 expression ( n = 4, scale bar: left, 100 µm; middle and right, 50 µm). Data are presented as the mean ± SEM; * p

    Article Snippet: The following primary antibodies were used in the procedures: rat monoclonal anti-CXCR4 antibody (R & D Systems, MAB21651), rabbit monoclonal anti-KRT7 antibody (Abcam, ab181598; Cambridge, UK); rabbit monoclonal anti-CDH1 antibody (CST, 3195; Beverly, MA, USA), rabbit monoclonal anti-H2AK119ub antibody (CST, 8240); mouse monoclonal anti-CXCL12 antibody (Novus Biologicals, MAB350; Littleton, CO, USA), rabbit polyclonal anti-AQP5 antibody (Alomone Labs, AQP-005; Jerusalem, Israel), rabbit monoclonal anti-CASP3 antibody (CST, 9664), and rat monoclonal anti-KI67 antibody (ThermoFisher, 14-5698-82).

    Techniques: Staining, Immunostaining, Expressing

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

    Journal: Biomaterials

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

    doi: 10.1016/j.biomaterials.2019.119245

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

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

    Techniques: Derivative Assay, Ex Vivo, Staining

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

    Journal: American Journal of Respiratory Cell and Molecular Biology

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

    doi: 10.1165/rcmb.2016-0071OC

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

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

    Techniques: Expressing, Microarray, Generated, Isolation

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

    Journal: American Journal of Respiratory Cell and Molecular Biology

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

    doi: 10.1165/rcmb.2016-0071OC

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

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

    Techniques: Expressing, Double Staining

    Epithelial differentiation potential. ( a ) Each SG clone was allowed to form salisphere-like cell structure followed by salivary epithelial cell differentiation as described in the methods. Salivary epithelial differentiation was evaluated by immunofluorescence staining for AQP5 and α-amylase. Scale bars represent 20 μm. ( b ) Expression of salivary epithelial differentiation-associated molecular markers including AMY1A , AQP5 , and BHLHA15 was examined by semi-quantitative RT-PCR. ( c ) To assess the multiple epithelial differentiation potential of the SG clones, each clonal population was subject to in vitro hepatogenic differentiation. The differentiation was evaluated by PAS staining. ( d ) Expression of hepatocyte differentiation markers including ALB and TJP1 was analyzed by RT-PCR.

    Journal: Scientific Reports

    Article Title: Single Cell Clones Purified from Human Parotid Glands Display Features of Multipotent Epitheliomesenchymal Stem Cells

    doi: 10.1038/srep36303

    Figure Lengend Snippet: Epithelial differentiation potential. ( a ) Each SG clone was allowed to form salisphere-like cell structure followed by salivary epithelial cell differentiation as described in the methods. Salivary epithelial differentiation was evaluated by immunofluorescence staining for AQP5 and α-amylase. Scale bars represent 20 μm. ( b ) Expression of salivary epithelial differentiation-associated molecular markers including AMY1A , AQP5 , and BHLHA15 was examined by semi-quantitative RT-PCR. ( c ) To assess the multiple epithelial differentiation potential of the SG clones, each clonal population was subject to in vitro hepatogenic differentiation. The differentiation was evaluated by PAS staining. ( d ) Expression of hepatocyte differentiation markers including ALB and TJP1 was analyzed by RT-PCR.

    Article Snippet: At the end of differentiation, cells were immunostained with anti-AQP5 antibody (Alomone Labs, Jerusalem, Israel) and anti-α-amylase antibody (Santa Cruz, CA, USA) to confirm that they were SG acinar cells.

    Techniques: Cell Differentiation, Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Clone Assay, In Vitro, Reverse Transcription Polymerase Chain Reaction

    Analysis of multilineage cell marker expression. Flow cytometric analysis was conducted to monitor the marker expression of clonal SG cells. Numbers above bracketed lines indicate the percentage of the cell population. Similar to MSCs, clonal SG cells commonly expressed MSC markers, but did not express hematopoietic/endothelial markers. Clonal SG cells were also positive for Oct4, but negative for Sox2. Basal expression of AQP5 was not detected and samples were negative for CD24, a recently suggested salivary progenitor marker. Blue line, each marker indicated; red line, isotype-matched control antibody.

    Journal: Scientific Reports

    Article Title: Single Cell Clones Purified from Human Parotid Glands Display Features of Multipotent Epitheliomesenchymal Stem Cells

    doi: 10.1038/srep36303

    Figure Lengend Snippet: Analysis of multilineage cell marker expression. Flow cytometric analysis was conducted to monitor the marker expression of clonal SG cells. Numbers above bracketed lines indicate the percentage of the cell population. Similar to MSCs, clonal SG cells commonly expressed MSC markers, but did not express hematopoietic/endothelial markers. Clonal SG cells were also positive for Oct4, but negative for Sox2. Basal expression of AQP5 was not detected and samples were negative for CD24, a recently suggested salivary progenitor marker. Blue line, each marker indicated; red line, isotype-matched control antibody.

    Article Snippet: At the end of differentiation, cells were immunostained with anti-AQP5 antibody (Alomone Labs, Jerusalem, Israel) and anti-α-amylase antibody (Santa Cruz, CA, USA) to confirm that they were SG acinar cells.

    Techniques: Marker, Expressing, Flow Cytometry

    Temporal expression patterns of some genes during salivary epithelial differentiation. As a representative GSC, clone C1 was subject to salivary epithelial differentiation for up to 14 days. RNA samples were isolated at the indicated time points. Q-PCR was conducted to examine the temporal expression patterns of genes including AMY, AQP5, CK5, CD24, CD90, CD49f, LGR5, and LGR6. Results are presented as the means ± SEM. One-way ANOVA and Tukey’s post hoc multiple comparison test were performed. *, vs. Day 0; #, vs. Day 1; $, vs. Day 7. (*p

    Journal: Scientific Reports

    Article Title: Single Cell Clones Purified from Human Parotid Glands Display Features of Multipotent Epitheliomesenchymal Stem Cells

    doi: 10.1038/srep36303

    Figure Lengend Snippet: Temporal expression patterns of some genes during salivary epithelial differentiation. As a representative GSC, clone C1 was subject to salivary epithelial differentiation for up to 14 days. RNA samples were isolated at the indicated time points. Q-PCR was conducted to examine the temporal expression patterns of genes including AMY, AQP5, CK5, CD24, CD90, CD49f, LGR5, and LGR6. Results are presented as the means ± SEM. One-way ANOVA and Tukey’s post hoc multiple comparison test were performed. *, vs. Day 0; #, vs. Day 1; $, vs. Day 7. (*p

    Article Snippet: At the end of differentiation, cells were immunostained with anti-AQP5 antibody (Alomone Labs, Jerusalem, Israel) and anti-α-amylase antibody (Santa Cruz, CA, USA) to confirm that they were SG acinar cells.

    Techniques: Expressing, Isolation, Polymerase Chain Reaction