aqp5  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs aqp5
    ATG5 exacerbates duct ligation-induced acinar cell atrophy. ( a ) Duct ligation induces acinar cell atrophy. Frozen sections of L3 SMGs and their contralateral counterparts (Ctrl) from <t>mT/mG;Aqp5-Cre</t> mice were visualized with fluorescence microscopy. Red fluorescence protein (RFP) was detected ubiquitously in all cells of mT/mG mice except for Aqp5 promoter-driven Cre -expressing acinar cells, which were marked by green fluorescence protein (GFP). Number of GFP-labeled acinar cells was considerably less in L3 gland than control gland. Bar: 100 μ m. ( b ) Suppression of ligation-induced acinar cell death in Atg5 KO mice. L3 FFPE SMG sections from Atg5 WT and Atg5 KO mice were immunostained for acinar-specific AQP5 protein. Number of AQP5-stained acinar cells was considerably less in L3 SMGs of Atg5 WT than that of Atg5 KO mice. Bar: 50 μ m
    Aqp5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aqp5/product/Alomone Labs
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    aqp5 - by Bioz Stars, 2022-08
    94/100 stars

    Images

    1) Product Images from "Dynamic involvement of ATG5 in cellular stress responses"

    Article Title: Dynamic involvement of ATG5 in cellular stress responses

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2014.428

    ATG5 exacerbates duct ligation-induced acinar cell atrophy. ( a ) Duct ligation induces acinar cell atrophy. Frozen sections of L3 SMGs and their contralateral counterparts (Ctrl) from mT/mG;Aqp5-Cre mice were visualized with fluorescence microscopy. Red fluorescence protein (RFP) was detected ubiquitously in all cells of mT/mG mice except for Aqp5 promoter-driven Cre -expressing acinar cells, which were marked by green fluorescence protein (GFP). Number of GFP-labeled acinar cells was considerably less in L3 gland than control gland. Bar: 100 μ m. ( b ) Suppression of ligation-induced acinar cell death in Atg5 KO mice. L3 FFPE SMG sections from Atg5 WT and Atg5 KO mice were immunostained for acinar-specific AQP5 protein. Number of AQP5-stained acinar cells was considerably less in L3 SMGs of Atg5 WT than that of Atg5 KO mice. Bar: 50 μ m
    Figure Legend Snippet: ATG5 exacerbates duct ligation-induced acinar cell atrophy. ( a ) Duct ligation induces acinar cell atrophy. Frozen sections of L3 SMGs and their contralateral counterparts (Ctrl) from mT/mG;Aqp5-Cre mice were visualized with fluorescence microscopy. Red fluorescence protein (RFP) was detected ubiquitously in all cells of mT/mG mice except for Aqp5 promoter-driven Cre -expressing acinar cells, which were marked by green fluorescence protein (GFP). Number of GFP-labeled acinar cells was considerably less in L3 gland than control gland. Bar: 100 μ m. ( b ) Suppression of ligation-induced acinar cell death in Atg5 KO mice. L3 FFPE SMG sections from Atg5 WT and Atg5 KO mice were immunostained for acinar-specific AQP5 protein. Number of AQP5-stained acinar cells was considerably less in L3 SMGs of Atg5 WT than that of Atg5 KO mice. Bar: 50 μ m

    Techniques Used: Ligation, Mouse Assay, Fluorescence, Microscopy, Expressing, Labeling, Formalin-fixed Paraffin-Embedded, Staining

    Elevated basal expression of proinflammatory cytokines genes in Atg5 -knockout SMGs. ( a ) Schematic diagram of generation of experimental mice. Mice developed a spontaneous heterologous Atg5 deletion after five generations of crossing between Atg5 f/f and Atg5 f/f ; Aqp5 -Cre mice, resulting in Atg5 f/− ; Aqp5 -Cre mice. Atg5 f/+ ; Aqp5 -Cre ( Atg5 WT ) and Atg5 f/− ; Aqp5 -Cre ( Atg5 KO ) mice were used in studies herein. ( b ) Immunohistochemical analyses show decreased ATG5 protein in both SMG granular convoluted ducts (GCDs; labeled D) and acinar cells (labeled A) of Atg5 KO mice, compared with that of Atg5 WT mice. Bar: 100 μ m. ( c ) Correlation of decreased ATG5 expression and impaired MAP1LC3 lipidation in SMGs among different genotypes. Equal amounts of whole SMG lysates from two individual mouse of the indicated genotype were analyzed for ATG5 and MAP1LC3 levels by western blots. ATG5 expression was greatly reduced in the SMGs of Atg5 F/F than that in Atg5 +/+ mice. This hypomorphic phenotype in floxed mouse line has been reported previously in the loxP mouse gene targeting system. 18 +/+ , Atg5 +/+ ; +/− , Atg5 +/ − ; +/−C , Atg5 +/ − ; Aqp5-Cre ; F/F , Atg5 F/F ; F/+ , Atg5 F/+ ; F/+C , Atg5 F/+ ; Aqp5-Cre (or Atg5 WT ); F/− , Atg5 F/ − ; F/−C , Atg5 F/ − ; Aqp5-Cre (or Atg5 KO ). Relative ATG5 levels were determined by setting average level of ATG5 in Atg5 +/+ mice as 1. ( d ) Quantitative RT-PCR analyses show elevated basal expression of selected proinflammatory cytokine genes in SMGs from Atg5 KO mice. Respective mean expression level of the indicated message from SMGs of Atg5 WT mice was set as 1. Results are shown as mean±S.D.; N =3; * P
    Figure Legend Snippet: Elevated basal expression of proinflammatory cytokines genes in Atg5 -knockout SMGs. ( a ) Schematic diagram of generation of experimental mice. Mice developed a spontaneous heterologous Atg5 deletion after five generations of crossing between Atg5 f/f and Atg5 f/f ; Aqp5 -Cre mice, resulting in Atg5 f/− ; Aqp5 -Cre mice. Atg5 f/+ ; Aqp5 -Cre ( Atg5 WT ) and Atg5 f/− ; Aqp5 -Cre ( Atg5 KO ) mice were used in studies herein. ( b ) Immunohistochemical analyses show decreased ATG5 protein in both SMG granular convoluted ducts (GCDs; labeled D) and acinar cells (labeled A) of Atg5 KO mice, compared with that of Atg5 WT mice. Bar: 100 μ m. ( c ) Correlation of decreased ATG5 expression and impaired MAP1LC3 lipidation in SMGs among different genotypes. Equal amounts of whole SMG lysates from two individual mouse of the indicated genotype were analyzed for ATG5 and MAP1LC3 levels by western blots. ATG5 expression was greatly reduced in the SMGs of Atg5 F/F than that in Atg5 +/+ mice. This hypomorphic phenotype in floxed mouse line has been reported previously in the loxP mouse gene targeting system. 18 +/+ , Atg5 +/+ ; +/− , Atg5 +/ − ; +/−C , Atg5 +/ − ; Aqp5-Cre ; F/F , Atg5 F/F ; F/+ , Atg5 F/+ ; F/+C , Atg5 F/+ ; Aqp5-Cre (or Atg5 WT ); F/− , Atg5 F/ − ; F/−C , Atg5 F/ − ; Aqp5-Cre (or Atg5 KO ). Relative ATG5 levels were determined by setting average level of ATG5 in Atg5 +/+ mice as 1. ( d ) Quantitative RT-PCR analyses show elevated basal expression of selected proinflammatory cytokine genes in SMGs from Atg5 KO mice. Respective mean expression level of the indicated message from SMGs of Atg5 WT mice was set as 1. Results are shown as mean±S.D.; N =3; * P

    Techniques Used: Expressing, Knock-Out, Mouse Assay, Immunohistochemistry, Labeling, Western Blot, Quantitative RT-PCR

    2) Product Images from "Stem cell properties of human clonal salivary gland stem cells are enhanced by three-dimensional priming culture in nanofibrous microwells"

    Article Title: Stem cell properties of human clonal salivary gland stem cells are enhanced by three-dimensional priming culture in nanofibrous microwells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-0829-x

    Enhanced differentiation potential of 3D-assembled SGSC spheroids. a Transcription levels of salivary acinar ( AMY1A and AQP5 ) and stem cell-related ( POU5F1 and THY1 ) markers compared by qPCR at 1, 3, 5, and 7 days between 2D monolayer-cultured SGSCs (SGSCs 2D ) and 3D spheroid-derived SGSCs (SGSCs 3D ) culture. Data from three independent experiments analyzed and presented as mean ± SEM ( n = 3). Two-way ANOVA, Bonferroni’s post hoc test. *Compared with Day 1; # compared with SGSCs 2D in each group. b Light microscope images of SGSCs after differentiation. Scale bars represent 400 μm. c Differentiation capacity determined by measuring average number of acinus-like organoids per plate after plating same number of cells. Data from five independent experiments analyzed and presented as mean ± SEM ( n = 5). One-way ANOVA; Tukey’s post hoc test. *Compared with monolayer-cultured salivary gland-resident stem cells (SGSCs 2D ). d Immunofluorescent images representing salivary acinar markers α-amylase (red) and AQP5 (green), tight junction protein TJP1 (green), and adherence protein E-cadherin (red). Scale bars represent 20 μm. e mRNA levels of SG acinar cell markers (A MY1A and AQP5 ), tight junction gene ( TJP1 ), and intercellular adherence gene ( CDH1 ) determined by real-time PCR in SGSC 2D and SGSC 3D cultures after differentiation. All qPCR measurements performed in triplicate. f Protein levels of α-amylase, AQP5, TJP1, and E-cadherin determined by western blotting in SGSC 2D and SGSC 3D cultures after differentiation. ** P
    Figure Legend Snippet: Enhanced differentiation potential of 3D-assembled SGSC spheroids. a Transcription levels of salivary acinar ( AMY1A and AQP5 ) and stem cell-related ( POU5F1 and THY1 ) markers compared by qPCR at 1, 3, 5, and 7 days between 2D monolayer-cultured SGSCs (SGSCs 2D ) and 3D spheroid-derived SGSCs (SGSCs 3D ) culture. Data from three independent experiments analyzed and presented as mean ± SEM ( n = 3). Two-way ANOVA, Bonferroni’s post hoc test. *Compared with Day 1; # compared with SGSCs 2D in each group. b Light microscope images of SGSCs after differentiation. Scale bars represent 400 μm. c Differentiation capacity determined by measuring average number of acinus-like organoids per plate after plating same number of cells. Data from five independent experiments analyzed and presented as mean ± SEM ( n = 5). One-way ANOVA; Tukey’s post hoc test. *Compared with monolayer-cultured salivary gland-resident stem cells (SGSCs 2D ). d Immunofluorescent images representing salivary acinar markers α-amylase (red) and AQP5 (green), tight junction protein TJP1 (green), and adherence protein E-cadherin (red). Scale bars represent 20 μm. e mRNA levels of SG acinar cell markers (A MY1A and AQP5 ), tight junction gene ( TJP1 ), and intercellular adherence gene ( CDH1 ) determined by real-time PCR in SGSC 2D and SGSC 3D cultures after differentiation. All qPCR measurements performed in triplicate. f Protein levels of α-amylase, AQP5, TJP1, and E-cadherin determined by western blotting in SGSC 2D and SGSC 3D cultures after differentiation. ** P

    Techniques Used: Real-time Polymerase Chain Reaction, Cell Culture, Derivative Assay, Light Microscopy, Western Blot

    3) Product Images from "Mesenchymal Nuclear factor I B regulates cell proliferation and epithelial differentiation during lung maturation"

    Article Title: Mesenchymal Nuclear factor I B regulates cell proliferation and epithelial differentiation during lung maturation

    Journal: Developmental biology

    doi: 10.1016/j.ydbio.2011.04.002

    Loss of Nfib in mesenchyme affects type I and type II epithelial cell differentiation. E18.5 lung sections from Nfib flox/flox and Nfib flox/flox , D1-Cre embryos were stained for AQP5 (A, B), pro-SPC (C, D) and CC10 (E, F). AQP5, pro-SPC and Foxj1 transcript
    Figure Legend Snippet: Loss of Nfib in mesenchyme affects type I and type II epithelial cell differentiation. E18.5 lung sections from Nfib flox/flox and Nfib flox/flox , D1-Cre embryos were stained for AQP5 (A, B), pro-SPC (C, D) and CC10 (E, F). AQP5, pro-SPC and Foxj1 transcript

    Techniques Used: Cell Differentiation, Staining

    4) Product Images from "Biocompatible Tissue Scaffold Compliance Promotes Salivary Gland Morphogenesis and Differentiation"

    Article Title: Biocompatible Tissue Scaffold Compliance Promotes Salivary Gland Morphogenesis and Differentiation

    Journal: Tissue Engineering. Part A

    doi: 10.1089/ten.tea.2013.0515

    The aberrant development of salivary gland organ explants grown on a stiff substrate can be rescued with a compliant substrate. (A) Mechanical rescue of branching morphogenesis. Brightfield images show representative E13 salivary glands grown at 0.48 and 19.66 kPa for 96 h with disruption of both bud morphology and number that is evident at 19.66 kPa. When transferred to a 0.48 kPa PA gel after 72 h culture on a 19.66 kPa gel, the glands regain a normal bud morphology and number similar to glands cultured on 0.48 kPa PA gels continuously. The white star indicates a transferred gland; scale bar=500 μm. (B–D) Mechanical rescue of epithelial differentiation marker expression levels. (B) Western analysis indicates a decrease in both AQP5 and SM α-actin with increasing stiffness. There is a near-complete rescue of both AQP5 and SM α-actin protein levels when glands are transferred from 19.66 kPa gel to the compliant 0.48 kPa gel. (C, D) Quantification of the western analysis of AQP5 (C) and SM α-actin (D) , normalized to GAPDH and graphed as the fold change relative to the 0.48 kPa pixel density. A one-way ANOVA test with Bonferroni post-tests was applied for each protein ( n =4 experiments, ** p
    Figure Legend Snippet: The aberrant development of salivary gland organ explants grown on a stiff substrate can be rescued with a compliant substrate. (A) Mechanical rescue of branching morphogenesis. Brightfield images show representative E13 salivary glands grown at 0.48 and 19.66 kPa for 96 h with disruption of both bud morphology and number that is evident at 19.66 kPa. When transferred to a 0.48 kPa PA gel after 72 h culture on a 19.66 kPa gel, the glands regain a normal bud morphology and number similar to glands cultured on 0.48 kPa PA gels continuously. The white star indicates a transferred gland; scale bar=500 μm. (B–D) Mechanical rescue of epithelial differentiation marker expression levels. (B) Western analysis indicates a decrease in both AQP5 and SM α-actin with increasing stiffness. There is a near-complete rescue of both AQP5 and SM α-actin protein levels when glands are transferred from 19.66 kPa gel to the compliant 0.48 kPa gel. (C, D) Quantification of the western analysis of AQP5 (C) and SM α-actin (D) , normalized to GAPDH and graphed as the fold change relative to the 0.48 kPa pixel density. A one-way ANOVA test with Bonferroni post-tests was applied for each protein ( n =4 experiments, ** p

    Techniques Used: Cell Culture, Marker, Expressing, Western Blot

    Salivary gland proacinar morphology and epithelial differentiation are physiologically advanced by a compliant substrate and disrupted by aberrant stiffness. (A, B) Expression levels of differentiation markers. (A) Western analysis indicates a decrease in protein levels of both aquaporin 5 (AQP5) and SM α-actin with increasing stiffness. (B) Quantification of western analysis for AQP5 and SM α-actin protein levels, graphed as the relative pixel density of each band relative to the GAPDH control. A two-way ANOVA test with Bonferroni post-tests was applied at each time point ( n =6 experiments, * p
    Figure Legend Snippet: Salivary gland proacinar morphology and epithelial differentiation are physiologically advanced by a compliant substrate and disrupted by aberrant stiffness. (A, B) Expression levels of differentiation markers. (A) Western analysis indicates a decrease in protein levels of both aquaporin 5 (AQP5) and SM α-actin with increasing stiffness. (B) Quantification of western analysis for AQP5 and SM α-actin protein levels, graphed as the relative pixel density of each band relative to the GAPDH control. A two-way ANOVA test with Bonferroni post-tests was applied at each time point ( n =6 experiments, * p

    Techniques Used: Expressing, Western Blot

    The aberrant development of salivary glands grown on high-stiffness substrates is partially rescued with exogenous TGFβ1. (A–C) Chemical rescue of epithelial differentiation marker expression levels. (A) Western analysis indicates a decrease in AQP5 and SM α-actin with increasing stiffness that is partially rescued with exogenous TGFβ1 added to the culture media. (B, C) Quantification of western analysis to detect AQP5 and SM α-actin in response to TGFβ1, normalized to GAPDH and expressed as the fold change relative to the 0.48 kPa value. A one-way ANOVA test with Bonferroni post-tests was applied for each protein ( n =5 experiments, * p
    Figure Legend Snippet: The aberrant development of salivary glands grown on high-stiffness substrates is partially rescued with exogenous TGFβ1. (A–C) Chemical rescue of epithelial differentiation marker expression levels. (A) Western analysis indicates a decrease in AQP5 and SM α-actin with increasing stiffness that is partially rescued with exogenous TGFβ1 added to the culture media. (B, C) Quantification of western analysis to detect AQP5 and SM α-actin in response to TGFβ1, normalized to GAPDH and expressed as the fold change relative to the 0.48 kPa value. A one-way ANOVA test with Bonferroni post-tests was applied for each protein ( n =5 experiments, * p

    Techniques Used: Marker, Expressing, Western Blot

    5) Product Images from "FGF2-dependent mesenchyme and laminin-111 are niche factors in salivary gland organoids"

    Article Title: FGF2-dependent mesenchyme and laminin-111 are niche factors in salivary gland organoids

    Journal: Journal of Cell Science

    doi: 10.1242/jcs.208728

    FGF2 promotes elaboration of complex 3D salivary organoids in basement membrane. (A) Schematic of initial condition for epithelial clusters cultured with growth factors. (B) E16 epithelial clusters were seeded on porous polycarbonate filters floating on simple medium containing FGF2 or EGF. (C) Schematic of epithelial clusters cultured in basement membrane extract with growth factors. (D) Epithelial clusters were cultured in Matrigel with FGF2 or EGF. ICC and confocal imaging revealed that after 7 days, the EpCAM + epithelium is partially maintained with an increase in vimentin + mesenchyme cells with growth factors, while epithelial clusters grown in Matrigel with FGF2 form complex EpCAM + epithelial organoids with budded structures expressing AQP5. Staining was for EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). (E) Quantitative analysis shows the mean diameter for the epithelial clusters with or without growth factors in Matrigel. (F) Organoids were analyzed for the basal cell marker K14, the apical ductal cell marker K7 and the progenitor cell marker Kit in epithelial clusters after 7 days embedded in Matrigel with FGF2. Staining was for AQP5 (proacinar/acinar cells, red), K14 (basal, cyan), K7 (ductal, cyan) and Kit (progenitor, cyan) with DAPI (nuclei, blue). ** P
    Figure Legend Snippet: FGF2 promotes elaboration of complex 3D salivary organoids in basement membrane. (A) Schematic of initial condition for epithelial clusters cultured with growth factors. (B) E16 epithelial clusters were seeded on porous polycarbonate filters floating on simple medium containing FGF2 or EGF. (C) Schematic of epithelial clusters cultured in basement membrane extract with growth factors. (D) Epithelial clusters were cultured in Matrigel with FGF2 or EGF. ICC and confocal imaging revealed that after 7 days, the EpCAM + epithelium is partially maintained with an increase in vimentin + mesenchyme cells with growth factors, while epithelial clusters grown in Matrigel with FGF2 form complex EpCAM + epithelial organoids with budded structures expressing AQP5. Staining was for EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). (E) Quantitative analysis shows the mean diameter for the epithelial clusters with or without growth factors in Matrigel. (F) Organoids were analyzed for the basal cell marker K14, the apical ductal cell marker K7 and the progenitor cell marker Kit in epithelial clusters after 7 days embedded in Matrigel with FGF2. Staining was for AQP5 (proacinar/acinar cells, red), K14 (basal, cyan), K7 (ductal, cyan) and Kit (progenitor, cyan) with DAPI (nuclei, blue). ** P

    Techniques Used: Cell Culture, Immunocytochemistry, Imaging, Expressing, Staining, Marker

    Primary embryonic mesenchyme supports salivary organoid formation with robust AQP5 expression in co-culture
    Figure Legend Snippet: Primary embryonic mesenchyme supports salivary organoid formation with robust AQP5 expression in co-culture

    Techniques Used: Expressing, Co-Culture Assay

    Soluble factors from primary salivary mesenchyme cannot substitute for mesenchyme cells to support AQP5 proacinar epithelial cells in culture. (A) Schematic of initial conditions for epithelial clusters cultured without direct contact with the mesenchyme when separated by a filter or when cultured with mesenchymal cell conditioned medium for 7 days. (B) E16 epithelial clusters grown in Matrigel on a polycarbonate filter were separated from primary mesenchyme cells, which were seeded on a coverslip below. ICC and confocal imaging revealed loss of AQP5 expression with partial preservation of EpCAM + epithelium after 7 days. (C) E16 epithelial clusters were grown in Matrigel or laminin-111 in the presence of concentrated conditioned medium (cm) collected from E16 primary salivary mesenchyme. The conditioned medium (D) partially preserved the EpCAM + epithelial phenotype in the presence of Matrigel (mat) or laminin (lam) but (E) poorly rescued AQP5 expression, as determined by staining for the markers EpCAM (epithelium, green) and AQP5 (proacinar/acinar cells, red) with DAPI (nuclei, blue). ** P
    Figure Legend Snippet: Soluble factors from primary salivary mesenchyme cannot substitute for mesenchyme cells to support AQP5 proacinar epithelial cells in culture. (A) Schematic of initial conditions for epithelial clusters cultured without direct contact with the mesenchyme when separated by a filter or when cultured with mesenchymal cell conditioned medium for 7 days. (B) E16 epithelial clusters grown in Matrigel on a polycarbonate filter were separated from primary mesenchyme cells, which were seeded on a coverslip below. ICC and confocal imaging revealed loss of AQP5 expression with partial preservation of EpCAM + epithelium after 7 days. (C) E16 epithelial clusters were grown in Matrigel or laminin-111 in the presence of concentrated conditioned medium (cm) collected from E16 primary salivary mesenchyme. The conditioned medium (D) partially preserved the EpCAM + epithelial phenotype in the presence of Matrigel (mat) or laminin (lam) but (E) poorly rescued AQP5 expression, as determined by staining for the markers EpCAM (epithelium, green) and AQP5 (proacinar/acinar cells, red) with DAPI (nuclei, blue). ** P

    Techniques Used: Cell Culture, Immunocytochemistry, Imaging, Expressing, Preserving, Laser Capture Microdissection, Staining

    Mesenchyme-dependent salivary organoid formation requires FGF2 expression by the mesenchyme. (A) E16 epithelial clusters were seeded on primary mesenchyme feeder layers with the fibroblast growth factor receptor (FGFR) or epidermal growth factor (EGFR) inhibitors SU5402 and AG1478, respectively, in simple medium and were examined by ICC and confocal imaging. Quantitative analysis revealed loss of (B) epithelial EpCAM and (C) proacinar AQP5 protein expression with FGFR inhibition but not EGFR inhibition. (D) E16 mesenchyme cells were infected with lentiviral constructs expressing non-targeting (NT) shRNA or FGF2-targeting shRNA. An ELISA using cell lysates demonstrated a decline in FGF2 protein levels in mesenchymal cells infected with shRNA. (E) E16 epithelium was co-cultured with mesenchyme following lentiviral treatment with NT or FGF2 shRNA for 7 days. ICC and confocal imaging revealed (F) a decline in the epithelial population and (G) loss of proacinar differentiation, as shown by staining for the markers EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). * P
    Figure Legend Snippet: Mesenchyme-dependent salivary organoid formation requires FGF2 expression by the mesenchyme. (A) E16 epithelial clusters were seeded on primary mesenchyme feeder layers with the fibroblast growth factor receptor (FGFR) or epidermal growth factor (EGFR) inhibitors SU5402 and AG1478, respectively, in simple medium and were examined by ICC and confocal imaging. Quantitative analysis revealed loss of (B) epithelial EpCAM and (C) proacinar AQP5 protein expression with FGFR inhibition but not EGFR inhibition. (D) E16 mesenchyme cells were infected with lentiviral constructs expressing non-targeting (NT) shRNA or FGF2-targeting shRNA. An ELISA using cell lysates demonstrated a decline in FGF2 protein levels in mesenchymal cells infected with shRNA. (E) E16 epithelium was co-cultured with mesenchyme following lentiviral treatment with NT or FGF2 shRNA for 7 days. ICC and confocal imaging revealed (F) a decline in the epithelial population and (G) loss of proacinar differentiation, as shown by staining for the markers EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). * P

    Techniques Used: Expressing, Immunocytochemistry, Imaging, Inhibition, Infection, Construct, shRNA, Enzyme-linked Immunosorbent Assay, Cell Culture, Staining

    Complex laminin and FGF2-dependent salivary proacinar organoids require mesenchymal cells. (A) Schematic demonstrating isolation and culture of enriched or purified E16 epithelial clusters. GF, growth factor. (B) E16 enriched epithelial clusters were seeded in laminin-111 on porous polycarbonate filters floating on simple medium containing either FGF2 or EGF. (C) Brightfield images show enriched epithelial clusters or purified epithelial clusters after mesenchyme depletion cultured in Matrigel with FGF2 for 7 days. (D,E) E16 enriched epithelial clusters or E16 purified epithelial clusters (-Mes) were seeded in Matrigel and grown with FGF2. Laminin-111, FGF2 and mesenchyme when together lead to AQP5 expression being retained in organoids, as shown by staining for the markers EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). Quantitative analysis demonstrates (F) EpCAM is variably preserved in all conditions, while (G) AQP5 is retained with FGF2 and basement membrane only when mesenchyme is also present. * P
    Figure Legend Snippet: Complex laminin and FGF2-dependent salivary proacinar organoids require mesenchymal cells. (A) Schematic demonstrating isolation and culture of enriched or purified E16 epithelial clusters. GF, growth factor. (B) E16 enriched epithelial clusters were seeded in laminin-111 on porous polycarbonate filters floating on simple medium containing either FGF2 or EGF. (C) Brightfield images show enriched epithelial clusters or purified epithelial clusters after mesenchyme depletion cultured in Matrigel with FGF2 for 7 days. (D,E) E16 enriched epithelial clusters or E16 purified epithelial clusters (-Mes) were seeded in Matrigel and grown with FGF2. Laminin-111, FGF2 and mesenchyme when together lead to AQP5 expression being retained in organoids, as shown by staining for the markers EpCAM (epithelium, green), AQP5 (proacinar/acinar cells, red) and vimentin (mesenchyme, cyan) with DAPI (nuclei, blue). Quantitative analysis demonstrates (F) EpCAM is variably preserved in all conditions, while (G) AQP5 is retained with FGF2 and basement membrane only when mesenchyme is also present. * P

    Techniques Used: Isolation, Purification, Cell Culture, Expressing, Staining

    6) 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

    7) Product Images from "Stem cell properties of human clonal salivary gland stem cells are enhanced by three-dimensional priming culture in nanofibrous microwells"

    Article Title: Stem cell properties of human clonal salivary gland stem cells are enhanced by three-dimensional priming culture in nanofibrous microwells

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-018-0829-x

    Enhanced differentiation potential of 3D-assembled SGSC spheroids. a Transcription levels of salivary acinar ( AMY1A and AQP5 ) and stem cell-related ( POU5F1 and THY1 ) markers compared by qPCR at 1, 3, 5, and 7 days between 2D monolayer-cultured SGSCs (SGSCs 2D ) and 3D spheroid-derived SGSCs (SGSCs 3D ) culture. Data from three independent experiments analyzed and presented as mean ± SEM ( n = 3). Two-way ANOVA, Bonferroni’s post hoc test. *Compared with Day 1; # compared with SGSCs 2D in each group. b Light microscope images of SGSCs after differentiation. Scale bars represent 400 μm. c Differentiation capacity determined by measuring average number of acinus-like organoids per plate after plating same number of cells. Data from five independent experiments analyzed and presented as mean ± SEM ( n = 5). One-way ANOVA; Tukey’s post hoc test. *Compared with monolayer-cultured salivary gland-resident stem cells (SGSCs 2D ). d Immunofluorescent images representing salivary acinar markers α-amylase (red) and AQP5 (green), tight junction protein TJP1 (green), and adherence protein E-cadherin (red). Scale bars represent 20 μm. e mRNA levels of SG acinar cell markers (A MY1A and AQP5 ), tight junction gene ( TJP1 ), and intercellular adherence gene ( CDH1 ) determined by real-time PCR in SGSC 2D and SGSC 3D cultures after differentiation. All qPCR measurements performed in triplicate. f Protein levels of α-amylase, AQP5, TJP1, and E-cadherin determined by western blotting in SGSC 2D and SGSC 3D cultures after differentiation. ** P
    Figure Legend Snippet: Enhanced differentiation potential of 3D-assembled SGSC spheroids. a Transcription levels of salivary acinar ( AMY1A and AQP5 ) and stem cell-related ( POU5F1 and THY1 ) markers compared by qPCR at 1, 3, 5, and 7 days between 2D monolayer-cultured SGSCs (SGSCs 2D ) and 3D spheroid-derived SGSCs (SGSCs 3D ) culture. Data from three independent experiments analyzed and presented as mean ± SEM ( n = 3). Two-way ANOVA, Bonferroni’s post hoc test. *Compared with Day 1; # compared with SGSCs 2D in each group. b Light microscope images of SGSCs after differentiation. Scale bars represent 400 μm. c Differentiation capacity determined by measuring average number of acinus-like organoids per plate after plating same number of cells. Data from five independent experiments analyzed and presented as mean ± SEM ( n = 5). One-way ANOVA; Tukey’s post hoc test. *Compared with monolayer-cultured salivary gland-resident stem cells (SGSCs 2D ). d Immunofluorescent images representing salivary acinar markers α-amylase (red) and AQP5 (green), tight junction protein TJP1 (green), and adherence protein E-cadherin (red). Scale bars represent 20 μm. e mRNA levels of SG acinar cell markers (A MY1A and AQP5 ), tight junction gene ( TJP1 ), and intercellular adherence gene ( CDH1 ) determined by real-time PCR in SGSC 2D and SGSC 3D cultures after differentiation. All qPCR measurements performed in triplicate. f Protein levels of α-amylase, AQP5, TJP1, and E-cadherin determined by western blotting in SGSC 2D and SGSC 3D cultures after differentiation. ** P

    Techniques Used: Real-time Polymerase Chain Reaction, Cell Culture, Derivative Assay, Light Microscopy, Western Blot

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    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
    Rabbit Polyclonal Anti Aqp5 Antibody, 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/rabbit polyclonal anti aqp5 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti aqp5 antibody - by Bioz Stars, 2022-08
    94/100 stars
      Buy from Supplier

    Image Search Results


    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

    AdHSP reduces the abundance of caspases 8 and 9 in 2CLP-induced lung injury. A. Immunoblotting of whole cytosolic extracts for Caspase 8 and 9. Abbreviations as in Fig. 1 . Upper panel : Representative autoradiogram of SDS-PAGE, 30 µg of cytosolic extract/lane. Primary rabbit polyclonal antibody to caspase 8. Secondary goat anti rabbit IgG. .Graphic representation of relative density of cytosolic caspase 8. Middle panel : Representative autoradiogram of SDS-PAGE, 30 µg of cytosolic extracts. Primary rabbit polyclonal antibody to caspase 9, secondary goat anti rabbit IgG. Graph - Graphic representation of relative density (mean +/− standard deviation) of cytosolic caspase 9. * = significantly different from 2CLPPBS and 2CLPAdGFP. Lower panels: Representative autoradiogram of SDS-PAGE, 30 µg of mitochondrial extract/lane. Primary mouse monoclonal antibody to Bcl2, secondary goat anti mouse IgG and primary mouse monoclonal antibody to COX IV, secondary goat anti mouse IgG. COX IV serves as mitochondrial loading control. B. Hsp70 in vivo interaction with apopotosomal Apaf-1. Representative autoradiograms. Samples were immunoprecipitated with a rabbit polyclonal antibody to Apaf-1 and subjected to SDS-PAGE. Upper panels: Immunoblotting with a primary rabbit polyclonal antibody to pro-caspase 9, secondary goat anti rabbit IgG. Middle panels: Immunoblotting with primary mouse monoclonal antibody to Hsp70, secondary goat anti mouse IgG. Lower panels: IgG detection IgG serves as loading control. 250 µg of cytosolic extracts obtained from TO, 2CLPPBS, 2CLPAdHSP or 2CLPAdGFP treated animals sacrificed 48 hrs after the induction of sepsis. C. Hsp70 in vitro, MLE-12 cells, interaction with apopotosomal Apaf-1. Representative autoradiograms. 250 µg of cytosolic extracts obtained from non treated MLE-12 cells (controls), stimulated with tumor necrosis factor (TNF) and treated with AdHSP or AdGFP. Samples were immunoprecipitated with a rabbit polyclonal antibody to Apaf-1 and subjected to SDS-PAGE. Upper panels: Immunoblotting with a primary rabbit polyclonal antibody to pro-caspase 9, secondary goat anti rabbit IgG. Middle panels: Immunoblotting with primary mouse monoclonal antibody to Hsp70, secondary goat anti mouse IgG. Lower panels: IgG serves as loading control. D. Apaf-1 – CARD dissociates from Pro-caspase-9 in the presence of Hsp70. Representative autoradiograms. 100 µg of cytosolic extracts obtained from 2CLPPBS and 2CLAdHSP treated animals, were immunoprecipitated with Pro-caspase-9 and further incubated with GST-Apaf-1 – CARD obtained from BL-21 cells, together with 5 mM ATP and 5 µg/ml human Cytochrome C for 5, 10, 20 and 30 minutes. Samples were subjected to SDS-PAGE, immunoblotted and the membranes were incubated with primary rabbit polyclonal antibody to Cleaved Caspase-9, secondary to goat anti rabbit IgG.

    Journal: PLoS ONE

    Article Title: Enhanced Hsp70 Expression Protects against Acute Lung Injury by Modulating Apoptotic Pathways

    doi: 10.1371/journal.pone.0026956

    Figure Lengend Snippet: AdHSP reduces the abundance of caspases 8 and 9 in 2CLP-induced lung injury. A. Immunoblotting of whole cytosolic extracts for Caspase 8 and 9. Abbreviations as in Fig. 1 . Upper panel : Representative autoradiogram of SDS-PAGE, 30 µg of cytosolic extract/lane. Primary rabbit polyclonal antibody to caspase 8. Secondary goat anti rabbit IgG. .Graphic representation of relative density of cytosolic caspase 8. Middle panel : Representative autoradiogram of SDS-PAGE, 30 µg of cytosolic extracts. Primary rabbit polyclonal antibody to caspase 9, secondary goat anti rabbit IgG. Graph - Graphic representation of relative density (mean +/− standard deviation) of cytosolic caspase 9. * = significantly different from 2CLPPBS and 2CLPAdGFP. Lower panels: Representative autoradiogram of SDS-PAGE, 30 µg of mitochondrial extract/lane. Primary mouse monoclonal antibody to Bcl2, secondary goat anti mouse IgG and primary mouse monoclonal antibody to COX IV, secondary goat anti mouse IgG. COX IV serves as mitochondrial loading control. B. Hsp70 in vivo interaction with apopotosomal Apaf-1. Representative autoradiograms. Samples were immunoprecipitated with a rabbit polyclonal antibody to Apaf-1 and subjected to SDS-PAGE. Upper panels: Immunoblotting with a primary rabbit polyclonal antibody to pro-caspase 9, secondary goat anti rabbit IgG. Middle panels: Immunoblotting with primary mouse monoclonal antibody to Hsp70, secondary goat anti mouse IgG. Lower panels: IgG detection IgG serves as loading control. 250 µg of cytosolic extracts obtained from TO, 2CLPPBS, 2CLPAdHSP or 2CLPAdGFP treated animals sacrificed 48 hrs after the induction of sepsis. C. Hsp70 in vitro, MLE-12 cells, interaction with apopotosomal Apaf-1. Representative autoradiograms. 250 µg of cytosolic extracts obtained from non treated MLE-12 cells (controls), stimulated with tumor necrosis factor (TNF) and treated with AdHSP or AdGFP. Samples were immunoprecipitated with a rabbit polyclonal antibody to Apaf-1 and subjected to SDS-PAGE. Upper panels: Immunoblotting with a primary rabbit polyclonal antibody to pro-caspase 9, secondary goat anti rabbit IgG. Middle panels: Immunoblotting with primary mouse monoclonal antibody to Hsp70, secondary goat anti mouse IgG. Lower panels: IgG serves as loading control. D. Apaf-1 – CARD dissociates from Pro-caspase-9 in the presence of Hsp70. Representative autoradiograms. 100 µg of cytosolic extracts obtained from 2CLPPBS and 2CLAdHSP treated animals, were immunoprecipitated with Pro-caspase-9 and further incubated with GST-Apaf-1 – CARD obtained from BL-21 cells, together with 5 mM ATP and 5 µg/ml human Cytochrome C for 5, 10, 20 and 30 minutes. Samples were subjected to SDS-PAGE, immunoblotted and the membranes were incubated with primary rabbit polyclonal antibody to Cleaved Caspase-9, secondary to goat anti rabbit IgG.

    Article Snippet: AQP5 was detected using a primary rabbit polyclonal antibody (Almone Inc., Jerusalem, Israel) diluted 1∶100 and a secondary anti rabbit IgG.

    Techniques: SDS Page, Standard Deviation, In Vivo, Immunoprecipitation, In Vitro, Incubation

    AdHSP alters interactions between Caspase 8, 9, 3 and Apaf-1. A and B: Representative Autoradiogram Demonstrating AdHSP treatment disrupts interaction between caspase 8 and caspase 9. Representative autoradiograms. 250 µg of cytosolic extracts were immunoprecipitated with rabbit polyclonal antibody to caspase 8 or caspase-9 and subjected to SDS-PAGE. Immunoblotting performed with either primary rabbit polyclonal antibodies to caspase 9 or caspase-8, secondary goat anti rabbit IgG. Lower panels: IgG detection. IgG serves as loading control. Abbreviations as in Figure 1 . C: Hsp70 disrupts caspases 3, 8 9 and Apaf-1 complexes. 250 µg of cytosolic extracts from lung tissue fractionated via column chromatography, eluted by molecular weight, immunoprecipitated with an antibody to caspase-9 and subjected to 9% SDS-PAGE. Molecular weight of each fraction (kDa) indicated at the top of the figure. Detecting antibodies (anti-caspase 9, anti-caspase 8, anti-pro-caspase 3, anti-Hsp70 and anti-Apaf-1,) noted to the left of the panels. Lower panel: IgG detection. IgG serves as loading control. Abbreviations as in Figure 1 .

    Journal: PLoS ONE

    Article Title: Enhanced Hsp70 Expression Protects against Acute Lung Injury by Modulating Apoptotic Pathways

    doi: 10.1371/journal.pone.0026956

    Figure Lengend Snippet: AdHSP alters interactions between Caspase 8, 9, 3 and Apaf-1. A and B: Representative Autoradiogram Demonstrating AdHSP treatment disrupts interaction between caspase 8 and caspase 9. Representative autoradiograms. 250 µg of cytosolic extracts were immunoprecipitated with rabbit polyclonal antibody to caspase 8 or caspase-9 and subjected to SDS-PAGE. Immunoblotting performed with either primary rabbit polyclonal antibodies to caspase 9 or caspase-8, secondary goat anti rabbit IgG. Lower panels: IgG detection. IgG serves as loading control. Abbreviations as in Figure 1 . C: Hsp70 disrupts caspases 3, 8 9 and Apaf-1 complexes. 250 µg of cytosolic extracts from lung tissue fractionated via column chromatography, eluted by molecular weight, immunoprecipitated with an antibody to caspase-9 and subjected to 9% SDS-PAGE. Molecular weight of each fraction (kDa) indicated at the top of the figure. Detecting antibodies (anti-caspase 9, anti-caspase 8, anti-pro-caspase 3, anti-Hsp70 and anti-Apaf-1,) noted to the left of the panels. Lower panel: IgG detection. IgG serves as loading control. Abbreviations as in Figure 1 .

    Article Snippet: AQP5 was detected using a primary rabbit polyclonal antibody (Almone Inc., Jerusalem, Israel) diluted 1∶100 and a secondary anti rabbit IgG.

    Techniques: Immunoprecipitation, SDS Page, Column Chromatography, Molecular Weight

    AdHSP prevents nuclear translocation of activated Caspase-3. A. Representative autoradiograms for pro-caspase 3 and activated (cleaved) Caspase 3. 30 µg of cytosolic (upper panels) and nuclear (lower panel) extracts were subjected to SDS-PAGE. Immunoblotting performed with primary rabbit polyclonal antibody to pro- caspase 3 and secondary goat anti rabbit IgG, primary goat antibody to β-actin and secondary donkey anti goat IgG, primary rabbit polyclonal antibody to active (cleaved) caspase-3 and secondary goat anti rabbit IgG, primary mouse monoclonal antibody to histone (H1) and secondary goat anti mouse IgG.. β-actin and histone serve as loading controls. B. Representative stained fixed tissue section depicting intra-nuclear staining for Caspase 3. Sections obtained from T0 control, 2CLPPBS and 2CLPHSP rats. Tissue isolated 48 hrs after the induction of sepsis. Upper panel : 40× magnifications. Black arrows indicate active caspase 3 stained nuclei. Lower panel : 100× magnification of upper panel. C. Caspase 3 Activity Assay. Graphic representation of relative caspase-3 enzymatic activity (mean +/− standard deviation) * = significantly different from Control and AdHSP+TNF.

    Journal: PLoS ONE

    Article Title: Enhanced Hsp70 Expression Protects against Acute Lung Injury by Modulating Apoptotic Pathways

    doi: 10.1371/journal.pone.0026956

    Figure Lengend Snippet: AdHSP prevents nuclear translocation of activated Caspase-3. A. Representative autoradiograms for pro-caspase 3 and activated (cleaved) Caspase 3. 30 µg of cytosolic (upper panels) and nuclear (lower panel) extracts were subjected to SDS-PAGE. Immunoblotting performed with primary rabbit polyclonal antibody to pro- caspase 3 and secondary goat anti rabbit IgG, primary goat antibody to β-actin and secondary donkey anti goat IgG, primary rabbit polyclonal antibody to active (cleaved) caspase-3 and secondary goat anti rabbit IgG, primary mouse monoclonal antibody to histone (H1) and secondary goat anti mouse IgG.. β-actin and histone serve as loading controls. B. Representative stained fixed tissue section depicting intra-nuclear staining for Caspase 3. Sections obtained from T0 control, 2CLPPBS and 2CLPHSP rats. Tissue isolated 48 hrs after the induction of sepsis. Upper panel : 40× magnifications. Black arrows indicate active caspase 3 stained nuclei. Lower panel : 100× magnification of upper panel. C. Caspase 3 Activity Assay. Graphic representation of relative caspase-3 enzymatic activity (mean +/− standard deviation) * = significantly different from Control and AdHSP+TNF.

    Article Snippet: AQP5 was detected using a primary rabbit polyclonal antibody (Almone Inc., Jerusalem, Israel) diluted 1∶100 and a secondary anti rabbit IgG.

    Techniques: Translocation Assay, SDS Page, Staining, Isolation, Caspase-3 Activity Assay, Activity Assay, Standard Deviation