rabbit anti β galactosidase  (Valiant)


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    Name:
    Anti ß galactosidase no cross anti E coli rabbit IgG fraction fluorescein conjugated
    Description:
    Product is fluorescein 5 isothiocyanate FITC Isomer I comjugated rabbit IgG fraction to ß galactosidase no cross to E coli and buffer salts
    Catalog Number:
    0856032
    Price:
    347.3
    Applications:
    Immunoassays, Protein Purification, Flow Cytometry (FACS) , ELISA, Immunohistochemistry (Paraffin), Immunoblot
    Size:
    2 mL
    Category:
    Life Sciences Antibodies Primary Antibodies
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    Structured Review

    Valiant rabbit anti β galactosidase
    Expression of Robo receptors in migrating IO and LRN neurons. A–C , Consecutive coronal sections of E12 mouse embryos hybridized with digoxygenin-labeled riboprobes for Robo1 ( A ), Robo2 ( B ), and Robo3 ( C ). Precerebellar neurons leaving the rhombic lip express the three mRNAs (arrowheads). D , E , Three micrometer coronal sections of E13 Robo1 −/− embryos. Punctate <t>β-galactosidase</t> immunoreactivity is detected both in Brn3.2-immunopositive IO neurons ( D , arrowheads) (3 μm confocal image) and in LRN neurons (visualized by Hoechst staining) ( E , arrowheads) (3 μm confocal image). F–H , Coronal sections of E13 Robo2 −/− embryos. Diffuse β-galactosidase immunoreactivity is detected in IO neurons (IO in F ) and the marginal stream of migrating LRN neurons ( F , arrowheads). β-Galactosidase is coexpressed with Brn3.2 in IO neurons ( G ) and in the stream of LRN neurons visualized with Hoechst ( H , arrowhead) (3 μm confocal image). Scale bars: A–C , 170 μm; D , 8 μm; E , 20 μm; F , 100 μm; G , 26 μm; H , 10 μm. XII, Hypoglossal motor nucleus.
    Product is fluorescein 5 isothiocyanate FITC Isomer I comjugated rabbit IgG fraction to ß galactosidase no cross to E coli and buffer salts
    https://www.bioz.com/result/rabbit anti β galactosidase/product/Valiant
    Average 94 stars, based on 30 article reviews
    Price from $9.99 to $1999.99
    rabbit anti β galactosidase - by Bioz Stars, 2021-02
    94/100 stars

    Images

    1) Product Images from "Molecular Mechanisms Controlling Midline Crossing by Precerebellar Neurons"

    Article Title: Molecular Mechanisms Controlling Midline Crossing by Precerebellar Neurons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0078-08.2008

    Expression of Robo receptors in migrating IO and LRN neurons. A–C , Consecutive coronal sections of E12 mouse embryos hybridized with digoxygenin-labeled riboprobes for Robo1 ( A ), Robo2 ( B ), and Robo3 ( C ). Precerebellar neurons leaving the rhombic lip express the three mRNAs (arrowheads). D , E , Three micrometer coronal sections of E13 Robo1 −/− embryos. Punctate β-galactosidase immunoreactivity is detected both in Brn3.2-immunopositive IO neurons ( D , arrowheads) (3 μm confocal image) and in LRN neurons (visualized by Hoechst staining) ( E , arrowheads) (3 μm confocal image). F–H , Coronal sections of E13 Robo2 −/− embryos. Diffuse β-galactosidase immunoreactivity is detected in IO neurons (IO in F ) and the marginal stream of migrating LRN neurons ( F , arrowheads). β-Galactosidase is coexpressed with Brn3.2 in IO neurons ( G ) and in the stream of LRN neurons visualized with Hoechst ( H , arrowhead) (3 μm confocal image). Scale bars: A–C , 170 μm; D , 8 μm; E , 20 μm; F , 100 μm; G , 26 μm; H , 10 μm. XII, Hypoglossal motor nucleus.
    Figure Legend Snippet: Expression of Robo receptors in migrating IO and LRN neurons. A–C , Consecutive coronal sections of E12 mouse embryos hybridized with digoxygenin-labeled riboprobes for Robo1 ( A ), Robo2 ( B ), and Robo3 ( C ). Precerebellar neurons leaving the rhombic lip express the three mRNAs (arrowheads). D , E , Three micrometer coronal sections of E13 Robo1 −/− embryos. Punctate β-galactosidase immunoreactivity is detected both in Brn3.2-immunopositive IO neurons ( D , arrowheads) (3 μm confocal image) and in LRN neurons (visualized by Hoechst staining) ( E , arrowheads) (3 μm confocal image). F–H , Coronal sections of E13 Robo2 −/− embryos. Diffuse β-galactosidase immunoreactivity is detected in IO neurons (IO in F ) and the marginal stream of migrating LRN neurons ( F , arrowheads). β-Galactosidase is coexpressed with Brn3.2 in IO neurons ( G ) and in the stream of LRN neurons visualized with Hoechst ( H , arrowhead) (3 μm confocal image). Scale bars: A–C , 170 μm; D , 8 μm; E , 20 μm; F , 100 μm; G , 26 μm; H , 10 μm. XII, Hypoglossal motor nucleus.

    Techniques Used: Expressing, Labeling, Staining

    2) Product Images from "Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism"

    Article Title: Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.071761

    Dll1 , NICD and Hes1 expression in endoderm. ( A-D ) Optical sections of E7.5 to E10.5 Dll1 lacZ /+ embryos whole-mount stained for β-galactosidase, Cdh1, Pdx1, Neurog3 or Nkx6-1. The arrows in C point to Neurog3 + cells that co-express β-gal.
    Figure Legend Snippet: Dll1 , NICD and Hes1 expression in endoderm. ( A-D ) Optical sections of E7.5 to E10.5 Dll1 lacZ /+ embryos whole-mount stained for β-galactosidase, Cdh1, Pdx1, Neurog3 or Nkx6-1. The arrows in C point to Neurog3 + cells that co-express β-gal.

    Techniques Used: Expressing, Staining

    Ptf1a is required for Dll1 expression in MPCs. ( A-F ′) Image stack projections (A-C) and optical sections (D-E) of whole-mount stained E10.5 embryos of the indicated genotypes, stained for β-galactosidase indicating Dll1 expression, Pdx1
    Figure Legend Snippet: Ptf1a is required for Dll1 expression in MPCs. ( A-F ′) Image stack projections (A-C) and optical sections (D-E) of whole-mount stained E10.5 embryos of the indicated genotypes, stained for β-galactosidase indicating Dll1 expression, Pdx1

    Techniques Used: Expressing, Staining

    3) Product Images from "EphA4 deficient mice maintain astroglial-fibrotic scar formation after spinal cord injury"

    Article Title: EphA4 deficient mice maintain astroglial-fibrotic scar formation after spinal cord injury

    Journal: Experimental neurology

    doi: 10.1016/j.expneurol.2010.02.005

    β-gal reporter expression in EphA4−/− mice predominantly co-localizes with GFAP-positive cells in the spinal cord before and after dorsal hemisection SCI. GFAP (red) and β-gal (green) immunoreactivity from representative
    Figure Legend Snippet: β-gal reporter expression in EphA4−/− mice predominantly co-localizes with GFAP-positive cells in the spinal cord before and after dorsal hemisection SCI. GFAP (red) and β-gal (green) immunoreactivity from representative

    Techniques Used: Expressing, Mouse Assay

    A subset of β-gal expressing cells in EphA4−/− mice co-stain with NeuN in the spinal cord before and after SCI. NeuN (red) and β-gal (green) immunoreactivity from representative EphA4−/− (A, B) and wild
    Figure Legend Snippet: A subset of β-gal expressing cells in EphA4−/− mice co-stain with NeuN in the spinal cord before and after SCI. NeuN (red) and β-gal (green) immunoreactivity from representative EphA4−/− (A, B) and wild

    Techniques Used: Expressing, Mouse Assay, Staining

    β-gal reporter expression extensively co-localizes with neuronal marker NeuN in the sensorimotor cortex of adult mice. NeuN (red) and β-gal (green) immunoreactivity from representative EphA4−/− (A, B) and wild type control
    Figure Legend Snippet: β-gal reporter expression extensively co-localizes with neuronal marker NeuN in the sensorimotor cortex of adult mice. NeuN (red) and β-gal (green) immunoreactivity from representative EphA4−/− (A, B) and wild type control

    Techniques Used: Expressing, Marker, Mouse Assay

    4) Product Images from "Genome-wide identification of regulatory elements in Sertoli cells"

    Article Title: Genome-wide identification of regulatory elements in Sertoli cells

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.142554

    Transient transgenic analysis of a DHS unique to Sertoli cells identifies a new Sertoli cell enhancer upstream of Wt1 . (A) DNaseI-seq and H3K27ac ChIP-Seq data around the Wt1 locus. Gene names indicate the TSS of the transcript. Nearby genes are indicated in gray. DNaseI-Seq data (Parzen score) and peak calls (boxes above track) are in light blue; H3K27ac ChIP-seq data and peak calls are in dark blue. A box surrounds the DHS chosen for in vivo analysis. (B) Detailed view of the DHS showing the DNaseI-seq Parzen score, the smoothed base counts (light blue) and the H3K27ac data (dark blue). The light- and dark-blue bars indicate the DNaseI-seq and H3K27ac peaks, respectively. The black bar marks the region cloned upstream of an hsp68-LacZ reporter cassette. (C) An E13.5 testis from a transgenic embryo showed β-galactosidase expression ( TgWt1 , green) specifically in Sertoli cells. Sertoli cells were labeled by AMH immunostaining (red) and germ cells by CDH1 (blue). The confocal image was taken using a 20× objective. Scale bar: 100 µm. (D) Confocal image of the sample in C taken with a 40× objective. Scale bar: 12.5 µm. All three panels show germ cells (blue); the left panel shows Sertoli cells (red); the middle panel shows β-galactosidase expression (green); the right panel shows the merge (yellow indicates co-expression of β-galactosidase and AMH).
    Figure Legend Snippet: Transient transgenic analysis of a DHS unique to Sertoli cells identifies a new Sertoli cell enhancer upstream of Wt1 . (A) DNaseI-seq and H3K27ac ChIP-Seq data around the Wt1 locus. Gene names indicate the TSS of the transcript. Nearby genes are indicated in gray. DNaseI-Seq data (Parzen score) and peak calls (boxes above track) are in light blue; H3K27ac ChIP-seq data and peak calls are in dark blue. A box surrounds the DHS chosen for in vivo analysis. (B) Detailed view of the DHS showing the DNaseI-seq Parzen score, the smoothed base counts (light blue) and the H3K27ac data (dark blue). The light- and dark-blue bars indicate the DNaseI-seq and H3K27ac peaks, respectively. The black bar marks the region cloned upstream of an hsp68-LacZ reporter cassette. (C) An E13.5 testis from a transgenic embryo showed β-galactosidase expression ( TgWt1 , green) specifically in Sertoli cells. Sertoli cells were labeled by AMH immunostaining (red) and germ cells by CDH1 (blue). The confocal image was taken using a 20× objective. Scale bar: 100 µm. (D) Confocal image of the sample in C taken with a 40× objective. Scale bar: 12.5 µm. All three panels show germ cells (blue); the left panel shows Sertoli cells (red); the middle panel shows β-galactosidase expression (green); the right panel shows the merge (yellow indicates co-expression of β-galactosidase and AMH).

    Techniques Used: Transgenic Assay, Chromatin Immunoprecipitation, In Vivo, Clone Assay, Expressing, Labeling, Immunostaining

    5) Product Images from "Zipcode Binding Protein 1 Regulates the Development of Dendritic Arbors in Hippocampal Neurons"

    Article Title: Zipcode Binding Protein 1 Regulates the Development of Dendritic Arbors in Hippocampal Neurons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2387-10.2011

    β-Actin overexpression in hippocampal neurons with ZBP1 knockdown is sufficient for partial phenotype rescue. A , Micrographs of hippocampal neurons transfected on DIV7 with pEGFP-C1 together with pSuper vector as a control, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1 or EGFP-β-actin. Expression proceeded for 3 d. Neuronal morphology was visualized by staining for cotransfected β-gal. B–F , Mean number of primary dendrites ( B ), mean number of dendritic tips ( C ), dendritic branching index ( D ), mean total dendritic length ( E ), and Sholl analysis of transfected neurons ( F ). *** p
    Figure Legend Snippet: β-Actin overexpression in hippocampal neurons with ZBP1 knockdown is sufficient for partial phenotype rescue. A , Micrographs of hippocampal neurons transfected on DIV7 with pEGFP-C1 together with pSuper vector as a control, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1 or EGFP-β-actin. Expression proceeded for 3 d. Neuronal morphology was visualized by staining for cotransfected β-gal. B–F , Mean number of primary dendrites ( B ), mean number of dendritic tips ( C ), dendritic branching index ( D ), mean total dendritic length ( E ), and Sholl analysis of transfected neurons ( F ). *** p

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Expressing, Staining

    Overexpression of EGFP-ZBP1 in hippocampal neurons decreases dendrite branching. A , Representative micrographs of hippocampal neurons transfected on DIV7 for 5 d with control vector or EGFP-ZBP1. Neuronal morphology was visualized by staining for cotransfected β-gal. B , Sholl analysis of neurons transfected with EGFP-ZBP1 or control vector. The horizontal axis of the Sholl plot indicates the distance from the cell soma center. Scale bar, 20 μm.
    Figure Legend Snippet: Overexpression of EGFP-ZBP1 in hippocampal neurons decreases dendrite branching. A , Representative micrographs of hippocampal neurons transfected on DIV7 for 5 d with control vector or EGFP-ZBP1. Neuronal morphology was visualized by staining for cotransfected β-gal. B , Sholl analysis of neurons transfected with EGFP-ZBP1 or control vector. The horizontal axis of the Sholl plot indicates the distance from the cell soma center. Scale bar, 20 μm.

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Staining

    RNAi cleavage-resistant ZBP1 rescues ZBP1 knockdown phenotype in hippocampal neurons. A , Western blot analysis of the effect of ZBP1sh#1 and ZBP1sh#2 on the expression of cotransfected EGFP-ZBP1*1 in COS-7 cells. β-gal was cotransfected as a marker of even transfection. B , Representative micrographs of hippocampal neurons transfected on DIV7 with pSuper or pSuper-ZBP1sh#1 together with EGFP-ZBP1 or EGFP-ZBP1*1 for 3 d of expression. Neuronal morphology was visualized by staining for cotransfected β-gal. EGFP-ZBP1*1 was expressed regardless of the presence of ZBP1sh#1. C , Quantification of ZBP1 levels in neurons transfected with pSuper, ZBP1sh#1, and ZBP1sh#1 together with EGFP-ZBP1*1. Average immunofluorescence was measured in the cell body using MetaMorph software. D , Representative micrographs of hippocampal neurons transfected on DIV7 with control vector, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1 for 3 d of expression. Neuronal morphology was visualized by staining for cotransfected β-gal. E–I , Mean number of primary dendrites ( E ), mean number of dendritic tips ( F ), Sholl analysis ( G ), dendritic branching index ( H ), and mean total dendritic length of transfected neurons ( I ). *** p
    Figure Legend Snippet: RNAi cleavage-resistant ZBP1 rescues ZBP1 knockdown phenotype in hippocampal neurons. A , Western blot analysis of the effect of ZBP1sh#1 and ZBP1sh#2 on the expression of cotransfected EGFP-ZBP1*1 in COS-7 cells. β-gal was cotransfected as a marker of even transfection. B , Representative micrographs of hippocampal neurons transfected on DIV7 with pSuper or pSuper-ZBP1sh#1 together with EGFP-ZBP1 or EGFP-ZBP1*1 for 3 d of expression. Neuronal morphology was visualized by staining for cotransfected β-gal. EGFP-ZBP1*1 was expressed regardless of the presence of ZBP1sh#1. C , Quantification of ZBP1 levels in neurons transfected with pSuper, ZBP1sh#1, and ZBP1sh#1 together with EGFP-ZBP1*1. Average immunofluorescence was measured in the cell body using MetaMorph software. D , Representative micrographs of hippocampal neurons transfected on DIV7 with control vector, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1 for 3 d of expression. Neuronal morphology was visualized by staining for cotransfected β-gal. E–I , Mean number of primary dendrites ( E ), mean number of dendritic tips ( F ), Sholl analysis ( G ), dendritic branching index ( H ), and mean total dendritic length of transfected neurons ( I ). *** p

    Techniques Used: Western Blot, Expressing, Marker, Transfection, Staining, Immunofluorescence, Software, Plasmid Preparation

    Knockdown of ZBP1 in hippocampal neurons decreases dendritic branching. A , Representative images of hippocampal neurons transfected on DIV7 or DIV14 with pSuper, pSuper-ZBP1sh#1, or pSuper-ZBP1sh#2 for 3 d. Morphology of transfected cells was visualized by staining for cotransfected β-gal. B–F , Mean number of primary dendrites ( B ), mean total number of dendritic tips ( C ), dendritic branching index ( D ), mean total dendritic length ( E ), and Sholl analysis of neurons transfected on DIV7 ( F ). G , Representative images of hippocampal neurons transfected on DIV7 with pSuper and scrambled scZBP1sh and fixed 3 d later. Scale bar, 50 μm. H , Comparison of morphologies of dendritic arbors of neurons transfected with pSuper or scZBP1sh. The Sholl analysis revealed no significant difference. I–M , Mean number of primary dendrites ( I ), mean total number of dendritic tips ( J ), dendritic branching index ( K ), mean total dendritic length ( L ), and Sholl analysis of neurons transfected on DIV14 ( M ). *** p
    Figure Legend Snippet: Knockdown of ZBP1 in hippocampal neurons decreases dendritic branching. A , Representative images of hippocampal neurons transfected on DIV7 or DIV14 with pSuper, pSuper-ZBP1sh#1, or pSuper-ZBP1sh#2 for 3 d. Morphology of transfected cells was visualized by staining for cotransfected β-gal. B–F , Mean number of primary dendrites ( B ), mean total number of dendritic tips ( C ), dendritic branching index ( D ), mean total dendritic length ( E ), and Sholl analysis of neurons transfected on DIV7 ( F ). G , Representative images of hippocampal neurons transfected on DIV7 with pSuper and scrambled scZBP1sh and fixed 3 d later. Scale bar, 50 μm. H , Comparison of morphologies of dendritic arbors of neurons transfected with pSuper or scZBP1sh. The Sholl analysis revealed no significant difference. I–M , Mean number of primary dendrites ( I ), mean total number of dendritic tips ( J ), dendritic branching index ( K ), mean total dendritic length ( L ), and Sholl analysis of neurons transfected on DIV14 ( M ). *** p

    Techniques Used: Transfection, Staining

    Characterization of ZBP1 shRNAs. A–C , shRNA and scrambled RNA validation in COS-7 cell line. Cells were transfected with control or shRNA encoding plasmid together with either EGFP-ZBP1 or pEGFP-C1. Additionally, β-gal was cotransfected as a marker of even transfection. Both ZBP1sh#1 and ZBP1sh#2 strongly reduced EGFP-ZBP1 expression, whereas the scrambled version (scZBP1sh) did not. D–F , Verification of specificity of ZBP1shRNAs using qRT-PCR of cDNAs from neurons transfected on DIV0 with nucleofection with pSuper, ZBP1sh#1, ZBP1sh#2, and scrambled RNA. The graphs show the relative quantification of ZBP1 and two other members of the ZBP1 family: IMP2 and IMP3. * p
    Figure Legend Snippet: Characterization of ZBP1 shRNAs. A–C , shRNA and scrambled RNA validation in COS-7 cell line. Cells were transfected with control or shRNA encoding plasmid together with either EGFP-ZBP1 or pEGFP-C1. Additionally, β-gal was cotransfected as a marker of even transfection. Both ZBP1sh#1 and ZBP1sh#2 strongly reduced EGFP-ZBP1 expression, whereas the scrambled version (scZBP1sh) did not. D–F , Verification of specificity of ZBP1shRNAs using qRT-PCR of cDNAs from neurons transfected on DIV0 with nucleofection with pSuper, ZBP1sh#1, ZBP1sh#2, and scrambled RNA. The graphs show the relative quantification of ZBP1 and two other members of the ZBP1 family: IMP2 and IMP3. * p

    Techniques Used: shRNA, Transfection, Plasmid Preparation, Marker, Expressing, Quantitative RT-PCR

    ZBP1 mutant resistant to Src phosphorylation decreases dendritic branching when overexpressed and is not sufficient for phenotype rescue in neurons with endogenous ZBP1 knockdown. A , D , Representative micrographs of hippocampal neurons transfected on DIV7 for 5 d ( A ) or 3 d ( D ) with control vector, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F. Neuronal morphology was visualized by staining for cotransfected β-gal. B , C , E–G , Mean number of primary dendrites ( B ), mean total number of dendritic tips ( C ), dendritic branching index ( E ), total length of dendrites ( F ), and Sholl analysis of neurons overexpressing EGFP, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F for 5 d ( G ). H–J , Mean number of primary dendrites ( H ), mean total number of dendritic tips ( I ), and dendritic branching index of neurons overexpressing EGFP, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F for 3 d ( J ). *** p
    Figure Legend Snippet: ZBP1 mutant resistant to Src phosphorylation decreases dendritic branching when overexpressed and is not sufficient for phenotype rescue in neurons with endogenous ZBP1 knockdown. A , D , Representative micrographs of hippocampal neurons transfected on DIV7 for 5 d ( A ) or 3 d ( D ) with control vector, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F. Neuronal morphology was visualized by staining for cotransfected β-gal. B , C , E–G , Mean number of primary dendrites ( B ), mean total number of dendritic tips ( C ), dendritic branching index ( E ), total length of dendrites ( F ), and Sholl analysis of neurons overexpressing EGFP, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F for 5 d ( G ). H–J , Mean number of primary dendrites ( H ), mean total number of dendritic tips ( I ), and dendritic branching index of neurons overexpressing EGFP, EGFP-ZBP1*1, or EGFP-ZBP1*1-Y396F for 3 d ( J ). *** p

    Techniques Used: Mutagenesis, Transfection, Plasmid Preparation, Staining

    β-Actin mRNA and protein localization are disturbed in ZBP1-knockdown neurons. A , B , Representative micrographs of fluorescent β-actin in situ hybridization (red) in hippocampal neurons that were transfected on DIV7 with pSuper ( A ) or ZBP1sh#1 ( B ) for 36 h. Neuronal morphology is visualized by staining for cotransfected β-gal (green). Scale bar, 10 μm. A′ , A″ , B′ , and B″ show in greater magnification fragments of proximal ( A′ , B′ ) and distal ( A″ , B″ ) dendrites of transfected neurons. Scale bar, 10 μm. C , D , Hippocampal neurons were transfected at DIV7 for 48 h with pSuper, ZBP1sh#1, or ZBP1sh#2. Pictures show β-actin staining in proximal ( C ) and distal ( D ) dendrites of transfected cells. Scale bar, 10 μm. E , Mean number of β-actin mRNA granules per 10 μm of dendrite measured in proximal and distal dendrites (after branching). *** p
    Figure Legend Snippet: β-Actin mRNA and protein localization are disturbed in ZBP1-knockdown neurons. A , B , Representative micrographs of fluorescent β-actin in situ hybridization (red) in hippocampal neurons that were transfected on DIV7 with pSuper ( A ) or ZBP1sh#1 ( B ) for 36 h. Neuronal morphology is visualized by staining for cotransfected β-gal (green). Scale bar, 10 μm. A′ , A″ , B′ , and B″ show in greater magnification fragments of proximal ( A′ , B′ ) and distal ( A″ , B″ ) dendrites of transfected neurons. Scale bar, 10 μm. C , D , Hippocampal neurons were transfected at DIV7 for 48 h with pSuper, ZBP1sh#1, or ZBP1sh#2. Pictures show β-actin staining in proximal ( C ) and distal ( D ) dendrites of transfected cells. Scale bar, 10 μm. E , Mean number of β-actin mRNA granules per 10 μm of dendrite measured in proximal and distal dendrites (after branching). *** p

    Techniques Used: In Situ Hybridization, Transfection, Staining

    ZBP1 mutants with impaired RNA binding do not affect dendritic branching when overexpressed and are not sufficient for phenotype rescue in neurons with endogenous ZBP1 knockdown. A , Schematic representations of mutants used in this set of experiments. B , Representative micrographs of hippocampal neurons transfected on DIV7 with control vector, EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG. Neuronal morphology was visualized by staining for cotransfected β-gal. Scale bar, 20 μm. C–G , Mean number of primary dendrites ( C ), mean total number of dendritic tips ( D ), dendritic branching index ( E ), total length of dendrites ( F ), and Sholl analysis of neurons overexpressing EGFP, EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG ( G ). H , Representative micrographs of hippocampal neurons transfected on DIV7 with pEGFP-C1 together with empty pSuper vector as a control, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG. Expression proceeded for 3 d. Neuronal morphology was visualized by staining for cotransfected β-gal. Scale bar, 20 μm. I–K , Mean number of primary dendrites ( I ), mean total number of dendritic tips ( J ), and dendritic branching index of transfected neurons ( K ). *** p
    Figure Legend Snippet: ZBP1 mutants with impaired RNA binding do not affect dendritic branching when overexpressed and are not sufficient for phenotype rescue in neurons with endogenous ZBP1 knockdown. A , Schematic representations of mutants used in this set of experiments. B , Representative micrographs of hippocampal neurons transfected on DIV7 with control vector, EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG. Neuronal morphology was visualized by staining for cotransfected β-gal. Scale bar, 20 μm. C–G , Mean number of primary dendrites ( C ), mean total number of dendritic tips ( D ), dendritic branching index ( E ), total length of dendrites ( F ), and Sholl analysis of neurons overexpressing EGFP, EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG ( G ). H , Representative micrographs of hippocampal neurons transfected on DIV7 with pEGFP-C1 together with empty pSuper vector as a control, ZBP1sh#1, or ZBP1sh#1 together with EGFP-ZBP1*1, EGFP-ZBP1*1ΔC, or EGFP-ZBP1*1-GXXG. Expression proceeded for 3 d. Neuronal morphology was visualized by staining for cotransfected β-gal. Scale bar, 20 μm. I–K , Mean number of primary dendrites ( I ), mean total number of dendritic tips ( J ), and dendritic branching index of transfected neurons ( K ). *** p

    Techniques Used: RNA Binding Assay, Transfection, Plasmid Preparation, Staining, Expressing

    6) Product Images from "Col1a1+ perivascular cells in the brain are a source of retinoic acid following stroke"

    Article Title: Col1a1+ perivascular cells in the brain are a source of retinoic acid following stroke

    Journal: BMC Neuroscience

    doi: 10.1186/s12868-016-0284-5

    Assessment of in vivo RA signaling activity. Spatial RA activity assayed using RARE-hsp68-LacZ mouse 7 days post 60 min MCAO injury. Confocal, tile-scan image of β-galactosidase immunolabeling (β-gal, green , A ) with IB4 ( magenta , A ) at the level of the lesion (outlined with dashed line ). A′ and A″ indicate magnified images in A . Tile stitched confocal images of RA activity in non-ischemic hemispheres (β-gal, green , B , C ) and ischemic (β-gal, green , D – E ) with markers for neurons (NeuN, red, carets , B , D ), astrocytes (GFAP, red , carets , C , E ). Insets are magnified areas with dotted lines in D and E . Graphs depict quantification of NeuN+/β-gal+ neurons ( F ) and GFAP+/β-gal+ astrocytes ( G ) from non-ischemic and ischemic hemispheres 7 days following a 60 min MCAO (n ≥ 3 and bars represent SEM). Asterisks indicate statistically significant difference (p
    Figure Legend Snippet: Assessment of in vivo RA signaling activity. Spatial RA activity assayed using RARE-hsp68-LacZ mouse 7 days post 60 min MCAO injury. Confocal, tile-scan image of β-galactosidase immunolabeling (β-gal, green , A ) with IB4 ( magenta , A ) at the level of the lesion (outlined with dashed line ). A′ and A″ indicate magnified images in A . Tile stitched confocal images of RA activity in non-ischemic hemispheres (β-gal, green , B , C ) and ischemic (β-gal, green , D – E ) with markers for neurons (NeuN, red, carets , B , D ), astrocytes (GFAP, red , carets , C , E ). Insets are magnified areas with dotted lines in D and E . Graphs depict quantification of NeuN+/β-gal+ neurons ( F ) and GFAP+/β-gal+ astrocytes ( G ) from non-ischemic and ischemic hemispheres 7 days following a 60 min MCAO (n ≥ 3 and bars represent SEM). Asterisks indicate statistically significant difference (p

    Techniques Used: In Vivo, Activity Assay, Immunolabeling

    7) Product Images from "Ephrin-B3 reverse signaling through Grb4 and cytoskeletal regulators mediates axon pruning"

    Article Title: Ephrin-B3 reverse signaling through Grb4 and cytoskeletal regulators mediates axon pruning

    Journal: Nature neuroscience

    doi: 10.1038/nn.2254

    Defective hippocampal MF pruning in EB3 mutants. a , Confocal IF detects the EB3-β-gal fusion protein in calbinding-positive SPB (arrows) and IPB (arrowheads) MF axons extending from the DG towards the CA3 area in postnatal day 10 (PD10) and 10 week old adult (PW10) mice. The EphB2-β-gal fusion protein is expressed broadly above and underneath the MF bundles, but is not localized with calbindin in the MF axons. b and c , Anti-calbindin IF ( b ) and Timm stain ( c ) shows that IPB axons in 8–10 week old adult EB3 −/− mice are much longer than in WT littermates (distance between arrowheads). d , Quantification of the ratio of IPB length/length from hilus to curvature of CA3 area in EB3 −/− and WT littermates during postnatal development (n = 3–4 per group). e , Quantification of IPB length in EB3 −/− , EB3 lacZ/lacZ , and WT adult mice (n = 8–9 per group). Mean ± s.e.m. Scale bars: 300 μm in a , b and c (upper panels); 150 μm in b and c (bottom panels).
    Figure Legend Snippet: Defective hippocampal MF pruning in EB3 mutants. a , Confocal IF detects the EB3-β-gal fusion protein in calbinding-positive SPB (arrows) and IPB (arrowheads) MF axons extending from the DG towards the CA3 area in postnatal day 10 (PD10) and 10 week old adult (PW10) mice. The EphB2-β-gal fusion protein is expressed broadly above and underneath the MF bundles, but is not localized with calbindin in the MF axons. b and c , Anti-calbindin IF ( b ) and Timm stain ( c ) shows that IPB axons in 8–10 week old adult EB3 −/− mice are much longer than in WT littermates (distance between arrowheads). d , Quantification of the ratio of IPB length/length from hilus to curvature of CA3 area in EB3 −/− and WT littermates during postnatal development (n = 3–4 per group). e , Quantification of IPB length in EB3 −/− , EB3 lacZ/lacZ , and WT adult mice (n = 8–9 per group). Mean ± s.e.m. Scale bars: 300 μm in a , b and c (upper panels); 150 μm in b and c (bottom panels).

    Techniques Used: Mouse Assay, Staining

    8) Product Images from "Krüppel homolog 1 (Kr-h1) mediates juvenile hormone action during metamorphosis of Drosophila melanogaster"

    Article Title: Krüppel homolog 1 (Kr-h1) mediates juvenile hormone action during metamorphosis of Drosophila melanogaster

    Journal:

    doi: 10.1016/j.mod.2007.10.002

    Expression pattern of the β-galactosidase reporter of Kr-h1 activity in the enhancer-trap line cn 1 P{ry +t7.2 =PZ}Kr-h1 10642 /CyO; ry 506 after acetone (A, B) or JHM (C, D, E) treatment at puparium formation. Signal was detected by staining with 0.2%
    Figure Legend Snippet: Expression pattern of the β-galactosidase reporter of Kr-h1 activity in the enhancer-trap line cn 1 P{ry +t7.2 =PZ}Kr-h1 10642 /CyO; ry 506 after acetone (A, B) or JHM (C, D, E) treatment at puparium formation. Signal was detected by staining with 0.2%

    Techniques Used: Expressing, Activity Assay, Staining

    9) Product Images from "KCNK5 channels mostly expressed in cochlear outer sulcus cells are indispensable for hearing"

    Article Title: KCNK5 channels mostly expressed in cochlear outer sulcus cells are indispensable for hearing

    Journal: Nature Communications

    doi: 10.1038/ncomms9780

    Immunodetection of pendrin and/or β-galactosidase. In cryostat sections from adult cochleas, pendrin (yellow) appears in spiral prominence (Sp) and root cells (Rc) with a similar pattern of expression in Kcnk5 +/ − ( a ) and Kcnk5 −/− ( b ) mice at P45. DAPI (4′,6-diamidino-2-phenylindole dihydrochoride) staining of cell nuclei (blue). In adult Kcnk5 +/ − mouse cochlea ( c – k ), β-galactosidase (green) mainly found in Claudius' cells (Cc, c , d , i , j ), is also present in Rc ( e , k ), while pendrin (magenta) is localized in Rc ( c – h ). Merged images ( c – e ) show that pendrin and β-galactosidase only colocalize in Rc. Stacks of 23 confocal planes of 1.8 μm each in c , f and i , single confocal planes of 1.8 μm optic thickness in d , g and j and single confocal planes of 0.7 μm optic thickness in e , h and k . Scale bars, 50 μm ( a – c , f , i ); 20 μm ( d , e , g , h , j , k ).
    Figure Legend Snippet: Immunodetection of pendrin and/or β-galactosidase. In cryostat sections from adult cochleas, pendrin (yellow) appears in spiral prominence (Sp) and root cells (Rc) with a similar pattern of expression in Kcnk5 +/ − ( a ) and Kcnk5 −/− ( b ) mice at P45. DAPI (4′,6-diamidino-2-phenylindole dihydrochoride) staining of cell nuclei (blue). In adult Kcnk5 +/ − mouse cochlea ( c – k ), β-galactosidase (green) mainly found in Claudius' cells (Cc, c , d , i , j ), is also present in Rc ( e , k ), while pendrin (magenta) is localized in Rc ( c – h ). Merged images ( c – e ) show that pendrin and β-galactosidase only colocalize in Rc. Stacks of 23 confocal planes of 1.8 μm each in c , f and i , single confocal planes of 1.8 μm optic thickness in d , g and j and single confocal planes of 0.7 μm optic thickness in e , h and k . Scale bars, 50 μm ( a – c , f , i ); 20 μm ( d , e , g , h , j , k ).

    Techniques Used: Immunodetection, Expressing, Mouse Assay, Staining

    10) Product Images from "Epibranchial placode-derived neurons produce BDNF required for early sensory neuron development"

    Article Title: Epibranchial placode-derived neurons produce BDNF required for early sensory neuron development

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    doi: 10.1002/dvdy.22527

    Epibranchial placodes and their derivatives are labeled in Crect;ROSA mice (A–F) Horizontal sections of Crect;ROSA embryos. (A) In E9.5 Crect;ROSA embryos, β-galactosidase (green) is expressed in the surface ectoderm, including the epibranchial placodes (inset). At E11.5, β-galactosidase immunoreactivity is seen in placode-derived neurons of the (B) trigeminal (gV) and (C) geniculate (gVII) ganglia of Crect;ROSA embryos. β-galactosidase-IR cells are not present in the proximal portions of ganglia IX and X (D; PgIX and PgX) in Crect;ROSA embryos, but are present in the distal ganglia of IX and X (E; DgIX and DgX). (F) In the developing tongue of E11.5 Crect;ROSA embryos , most ingrowing neurites (arrows) are positive for neurofilament (red, F, F”), and a subset are immunopositive for β-galactosidase (green, F, F’). Fibers labeled exclusively by anti-neurofilament are indicated with an arrowhead. The tongue epithelium expresses β-galactosidase throughout at this stage (F, F’). (G–I) Horizontal sections of E11.5 Wnt1cre;ROSA embryo have β-galactosidase expression in neural crest-derived cells in the ganglia. (G) Neural crest derived β-galactosidase-IR cells (green) are detected on the surface of the geniculate ganglion (gVII ). Neural crest derived cells (green) are present throughout the proximal ganglia of IX and X (H; PgIX and PgX), and on the surface of the distal ganglia of IX and X (I; DgIX and DgX). Anterior is to the right. Blue: Hoechst stained nuclei. Scale bars are 20 µm in inset of A, 100 µm in A and F, and all others are 50 µm.
    Figure Legend Snippet: Epibranchial placodes and their derivatives are labeled in Crect;ROSA mice (A–F) Horizontal sections of Crect;ROSA embryos. (A) In E9.5 Crect;ROSA embryos, β-galactosidase (green) is expressed in the surface ectoderm, including the epibranchial placodes (inset). At E11.5, β-galactosidase immunoreactivity is seen in placode-derived neurons of the (B) trigeminal (gV) and (C) geniculate (gVII) ganglia of Crect;ROSA embryos. β-galactosidase-IR cells are not present in the proximal portions of ganglia IX and X (D; PgIX and PgX) in Crect;ROSA embryos, but are present in the distal ganglia of IX and X (E; DgIX and DgX). (F) In the developing tongue of E11.5 Crect;ROSA embryos , most ingrowing neurites (arrows) are positive for neurofilament (red, F, F”), and a subset are immunopositive for β-galactosidase (green, F, F’). Fibers labeled exclusively by anti-neurofilament are indicated with an arrowhead. The tongue epithelium expresses β-galactosidase throughout at this stage (F, F’). (G–I) Horizontal sections of E11.5 Wnt1cre;ROSA embryo have β-galactosidase expression in neural crest-derived cells in the ganglia. (G) Neural crest derived β-galactosidase-IR cells (green) are detected on the surface of the geniculate ganglion (gVII ). Neural crest derived cells (green) are present throughout the proximal ganglia of IX and X (H; PgIX and PgX), and on the surface of the distal ganglia of IX and X (I; DgIX and DgX). Anterior is to the right. Blue: Hoechst stained nuclei. Scale bars are 20 µm in inset of A, 100 µm in A and F, and all others are 50 µm.

    Techniques Used: Labeling, Mouse Assay, Derivative Assay, Expressing, Staining

    TrkB is expressed by neurons derived from epibranchial placodes but not by neural crest-derived cells At E11.5, full-length TrkB receptor immunoreactivity is present on most cells within the geniculate ganglion of Crect;ROSA (A–C) and Wnt1Cre;ROSA (D–F) mice. (A–C). Placodal cells express β-galactosidase (green, A, C) in Crect;ROSA embryos, and these cells are also TrkB immunoreactive (purple, B, C). Arrows indicate examples of double-labeled cells. (D–F). In Wnt1Cre;ROSA embryos, neural crest descendent cells are β-galactosidase immunoreactive (green), and comprise predominantly glia (arrowheads) with a small number of neurons (arrow) which do not express TrkB (purple, E, F). Anterior is up. Scale bar = 25µm.
    Figure Legend Snippet: TrkB is expressed by neurons derived from epibranchial placodes but not by neural crest-derived cells At E11.5, full-length TrkB receptor immunoreactivity is present on most cells within the geniculate ganglion of Crect;ROSA (A–C) and Wnt1Cre;ROSA (D–F) mice. (A–C). Placodal cells express β-galactosidase (green, A, C) in Crect;ROSA embryos, and these cells are also TrkB immunoreactive (purple, B, C). Arrows indicate examples of double-labeled cells. (D–F). In Wnt1Cre;ROSA embryos, neural crest descendent cells are β-galactosidase immunoreactive (green), and comprise predominantly glia (arrowheads) with a small number of neurons (arrow) which do not express TrkB (purple, E, F). Anterior is up. Scale bar = 25µm.

    Techniques Used: Derivative Assay, Mouse Assay, Labeling

    Epibranchial placodes contribute only neurons to the geniculate ganglia, while glial cells arise exclusively from neural crest (A–C). Horizontal sections of the geniculate ganglia in E11.5 Crect;ROSA embryos. (A) The early neuronal immunomarker Islet1/2 (purple, A, A’) labels many β–galactosidase labeled placode-derived cells (green) in Crect;ROSA embryos (arrows in A-A”). (B) An immunomarker of post-mitotic neurons, NeuN (purple, B, B’) also labels β–galactosidase –IR (green) placode-derived cells (arrows in B-B”). (C) Sox10, a glial cell immunomarker (purple, C, C’) does not co-localize with placodal cells labeled with β-galactosidase via Crect;ROSA (arrowheads in C-C” indicate examples of β-galactosidase immunoreactive cells that are not immunopositive for Sox10). (D–F) Horizontal sections of the geniculate ganglia in E11.5 Wnt1cre;ROSA embryos. (D,E) Most β-galactosidase labeled neural crest-derived cells (green) are negative for both neuronal markers (D; Islet1/2, and E; NeuN), although a few small-diameter neural crest cells are double immunopositive (arrow in E-E”). (F). Almost all Sox10-positive glial cells (purple, F-F”) were crest-derived, i.e., labeled with β-galactosidase (green, F-F”). Arrows in F-F” point out examples of double immunolabeled cells. Anterior is to the bottom right. Scale bar=25µm.
    Figure Legend Snippet: Epibranchial placodes contribute only neurons to the geniculate ganglia, while glial cells arise exclusively from neural crest (A–C). Horizontal sections of the geniculate ganglia in E11.5 Crect;ROSA embryos. (A) The early neuronal immunomarker Islet1/2 (purple, A, A’) labels many β–galactosidase labeled placode-derived cells (green) in Crect;ROSA embryos (arrows in A-A”). (B) An immunomarker of post-mitotic neurons, NeuN (purple, B, B’) also labels β–galactosidase –IR (green) placode-derived cells (arrows in B-B”). (C) Sox10, a glial cell immunomarker (purple, C, C’) does not co-localize with placodal cells labeled with β-galactosidase via Crect;ROSA (arrowheads in C-C” indicate examples of β-galactosidase immunoreactive cells that are not immunopositive for Sox10). (D–F) Horizontal sections of the geniculate ganglia in E11.5 Wnt1cre;ROSA embryos. (D,E) Most β-galactosidase labeled neural crest-derived cells (green) are negative for both neuronal markers (D; Islet1/2, and E; NeuN), although a few small-diameter neural crest cells are double immunopositive (arrow in E-E”). (F). Almost all Sox10-positive glial cells (purple, F-F”) were crest-derived, i.e., labeled with β-galactosidase (green, F-F”). Arrows in F-F” point out examples of double immunolabeled cells. Anterior is to the bottom right. Scale bar=25µm.

    Techniques Used: Labeling, Derivative Assay, Immunolabeling

    A subset of placode-derived geniculate ganglion cells produce BDNF, while neural crest-derived cells do not (A). In the geniculate ganglion of E11.5 Crect;bdnf lox/+ embryos, BDNF-producing placodal cells express BDNF-lacZ and are labeled with anti-β-galalactosidase (green, arrow). (B). In E11.5 Wnt1cre;bdnf lox/+ embryos, neural crest-derived cells in the geniculate ganglion (dashed outline) do not produce BDNF-lacZ and are not labeled via anti-β-galactosidase (green). However both gVIII and cells in the otic vesicle are β-galactosidase immunopositive. See text for details. Anterior is to the right. gVIII: acousticovestibular ganglion, ov: otic vesicle. gVII: geniculate ganglion is encircled with the dashed line. Anterior is to the right. Scale bar is 25µm.
    Figure Legend Snippet: A subset of placode-derived geniculate ganglion cells produce BDNF, while neural crest-derived cells do not (A). In the geniculate ganglion of E11.5 Crect;bdnf lox/+ embryos, BDNF-producing placodal cells express BDNF-lacZ and are labeled with anti-β-galalactosidase (green, arrow). (B). In E11.5 Wnt1cre;bdnf lox/+ embryos, neural crest-derived cells in the geniculate ganglion (dashed outline) do not produce BDNF-lacZ and are not labeled via anti-β-galactosidase (green). However both gVIII and cells in the otic vesicle are β-galactosidase immunopositive. See text for details. Anterior is to the right. gVIII: acousticovestibular ganglion, ov: otic vesicle. gVII: geniculate ganglion is encircled with the dashed line. Anterior is to the right. Scale bar is 25µm.

    Techniques Used: Derivative Assay, Labeling

    11) Product Images from "Nodal Expression in the Uterus of the Mouse Is Regulated by the Embryo and Correlates with Implantation 1"

    Article Title: Nodal Expression in the Uterus of the Mouse Is Regulated by the Embryo and Correlates with Implantation 1

    Journal: Biology of Reproduction

    doi: 10.1095/biolreprod.110.087239

    Immunohistochemical localization of NODAL in the mouse uterus throughout pregnancy. a ) Immunolocalization did not detect NODAL protein in the endometrium of nonpregnant (N.P.) females. b ) NODAL protein was observed within the glandular epithelium and lining the uterine lumen from Day 0.5 to 3.5 postcoitum (representative d3.5 shown). c ) On Day 8.5, NODAL was localized within the stromal and myometrial compartments exclusively on the antimesometrial aspect of the uterus. d ) By Day 12.5, NODAL protein was readily detectable in a distinct layer of the decidua parietalis on the lateral (shown) and mesometrial poles of the conceptus. e , g ) NODAL protein was also localized in spongiotrophoblast layer of the placenta during late pregnancy (Days 12.5 and 14.5 shown). f , h ) Although the β-galactosidase enzyme was consistently detectable in the decidua parietalis, spongiotrophoblast β-galactosidase was not observed in placentas of embryos that did not inherit the Nodal-lacZ allele, suggesting an extraembryonic origin. Bars = 50 μm. dp, decidua parietalis; la, labyrinth; sp, spongiotrophoblast.
    Figure Legend Snippet: Immunohistochemical localization of NODAL in the mouse uterus throughout pregnancy. a ) Immunolocalization did not detect NODAL protein in the endometrium of nonpregnant (N.P.) females. b ) NODAL protein was observed within the glandular epithelium and lining the uterine lumen from Day 0.5 to 3.5 postcoitum (representative d3.5 shown). c ) On Day 8.5, NODAL was localized within the stromal and myometrial compartments exclusively on the antimesometrial aspect of the uterus. d ) By Day 12.5, NODAL protein was readily detectable in a distinct layer of the decidua parietalis on the lateral (shown) and mesometrial poles of the conceptus. e , g ) NODAL protein was also localized in spongiotrophoblast layer of the placenta during late pregnancy (Days 12.5 and 14.5 shown). f , h ) Although the β-galactosidase enzyme was consistently detectable in the decidua parietalis, spongiotrophoblast β-galactosidase was not observed in placentas of embryos that did not inherit the Nodal-lacZ allele, suggesting an extraembryonic origin. Bars = 50 μm. dp, decidua parietalis; la, labyrinth; sp, spongiotrophoblast.

    Techniques Used: Immunohistochemistry

    Nodal-lacZ expression profile in the mouse uterus during late pregnancy. a ) Whole-mount staining on Day 10.5 was restricted to the mesometrial aspect (oriented right) of the uterus. b – c ) Transverse sectioning displayed prominent expression within a distinct layer of the decidua parietalis on the lateral and mesometrial surface of the conceptus. β-Galactosidase activity was readily detectable in the lateral ( d – f ) and mesometrial ( g – i ) decidua parietalis throughout the remainder of pregnancy as depicted in Day 12.5 and 14.5 representative samples. Original magnification ×4 ( b , e , h ) and ×10 ( c , f , i ).
    Figure Legend Snippet: Nodal-lacZ expression profile in the mouse uterus during late pregnancy. a ) Whole-mount staining on Day 10.5 was restricted to the mesometrial aspect (oriented right) of the uterus. b – c ) Transverse sectioning displayed prominent expression within a distinct layer of the decidua parietalis on the lateral and mesometrial surface of the conceptus. β-Galactosidase activity was readily detectable in the lateral ( d – f ) and mesometrial ( g – i ) decidua parietalis throughout the remainder of pregnancy as depicted in Day 12.5 and 14.5 representative samples. Original magnification ×4 ( b , e , h ) and ×10 ( c , f , i ).

    Techniques Used: Expressing, Staining, Activity Assay

    Nodal-lacZ expression profile in the mouse uterus during the peri-implantation period. a ) Whole-mount staining and ( b – c ) transverse sectioning of nonpregnant (N.P.) uteri did not reveal β-galactosidase activity at any point in the estrus cycle. d ) However, staining was readily detectable throughout the uterus after whole-mount staining from Day 0.5 to 3.5 postcoitum (representative d3.5 shown). e – f ) Transverse sectioning indicates expression was restricted to the glandular epithelium within the uterine endometrium. g ) On Day 4.5 postcoitum (d4.5), whole-mount staining generated a banding pattern along the proximal-distal axis of the uterine horn. h ) Serial transverse sections showed embryos undergoing implantation exclusively within the nonstained uterine bands, and ( i ) glandular epithelium at the implantation site was unstained, indicating Nodal expression was restricted to the sites between implantation crypts. Original magnification ×4 ( b , e ) and ×10 ( c , f , h , i ).
    Figure Legend Snippet: Nodal-lacZ expression profile in the mouse uterus during the peri-implantation period. a ) Whole-mount staining and ( b – c ) transverse sectioning of nonpregnant (N.P.) uteri did not reveal β-galactosidase activity at any point in the estrus cycle. d ) However, staining was readily detectable throughout the uterus after whole-mount staining from Day 0.5 to 3.5 postcoitum (representative d3.5 shown). e – f ) Transverse sectioning indicates expression was restricted to the glandular epithelium within the uterine endometrium. g ) On Day 4.5 postcoitum (d4.5), whole-mount staining generated a banding pattern along the proximal-distal axis of the uterine horn. h ) Serial transverse sections showed embryos undergoing implantation exclusively within the nonstained uterine bands, and ( i ) glandular epithelium at the implantation site was unstained, indicating Nodal expression was restricted to the sites between implantation crypts. Original magnification ×4 ( b , e ) and ×10 ( c , f , h , i ).

    Techniques Used: Expressing, Staining, Activity Assay, Generated

    Postimplantation Nodal-lacZ expression profile in the mouse uterus. a ) Whole-mount staining and ( b – c ) transverse sectioning of Day 6.5 uteri displayed significant β-galactosidase activity between decidua swellings within the glandular epithelium. d – f ) Within the implantation crypts, faint staining was observed in the stroma immediately adjacent the antimesometrial (oriented left) myometrium with decreasing β-galactosidase activity toward the central decidua. g ) By Day 8.5, increased staining was observed along the antimesometrial surface of the whole-mount uterus that was primarily localized within the endometrial stromal cells on the periphery of the zone of decidualization ( h – i ). j – l ) On Day 9.5, expression was observed within the stroma on the antimesometrial and lateral surfaces of the conceptus site. Original magnification ×2.5 ( e , h ), ×4 ( b , k ), ×10 ( l ), ×15 ( i ), and ×20 ( c , f ).
    Figure Legend Snippet: Postimplantation Nodal-lacZ expression profile in the mouse uterus. a ) Whole-mount staining and ( b – c ) transverse sectioning of Day 6.5 uteri displayed significant β-galactosidase activity between decidua swellings within the glandular epithelium. d – f ) Within the implantation crypts, faint staining was observed in the stroma immediately adjacent the antimesometrial (oriented left) myometrium with decreasing β-galactosidase activity toward the central decidua. g ) By Day 8.5, increased staining was observed along the antimesometrial surface of the whole-mount uterus that was primarily localized within the endometrial stromal cells on the periphery of the zone of decidualization ( h – i ). j – l ) On Day 9.5, expression was observed within the stroma on the antimesometrial and lateral surfaces of the conceptus site. Original magnification ×2.5 ( e , h ), ×4 ( b , k ), ×10 ( l ), ×15 ( i ), and ×20 ( c , f ).

    Techniques Used: Expressing, Staining, Activity Assay

    The embryo directs Nodal expression at the time of implantation. a ) Whole-mount staining revealed β-galactosidase activity throughout the entire uterus isolated from Day 0.5 to 3.5 pseudopregnant females (representative d3.5 shown). b ) In contrast to natural mating, Day 4.5 pseudopregnant uteri were devoid of staining. Embryo transfer of two ( c ) or six ( d ) blastocysts into a single uterine horn (*) restored the banding pattern on Day 4.5, and the number of nonstaining sites (arrows) correlated with the number of embryos introduced. Control horns were completely stained, indicating the embryo acts to maintain expression at the time of implantation, but inhibits Nodal expression at the implanting site.
    Figure Legend Snippet: The embryo directs Nodal expression at the time of implantation. a ) Whole-mount staining revealed β-galactosidase activity throughout the entire uterus isolated from Day 0.5 to 3.5 pseudopregnant females (representative d3.5 shown). b ) In contrast to natural mating, Day 4.5 pseudopregnant uteri were devoid of staining. Embryo transfer of two ( c ) or six ( d ) blastocysts into a single uterine horn (*) restored the banding pattern on Day 4.5, and the number of nonstaining sites (arrows) correlated with the number of embryos introduced. Control horns were completely stained, indicating the embryo acts to maintain expression at the time of implantation, but inhibits Nodal expression at the implanting site.

    Techniques Used: Expressing, Staining, Activity Assay, Isolation

    12) Product Images from "Pathogenic Stimulation of Intestinal Stem Cell response in Drosophila"

    Article Title: Pathogenic Stimulation of Intestinal Stem Cell response in Drosophila

    Journal: Journal of cellular physiology

    doi: 10.1002/jcp.21808

    Pathogen feeding does not alter the cell fate decision. Dissected guts from Su(H)-lacZ flies fed with the various agents as indicated were used for immunofluorescent staining. Delta staining (green) and β-galactosidase staining (red) were performed together on the guts. Representative confocal images are shown here. In control samples, the Delta-positive cells (A, arrow) and the Su(H)-lacZ positive cells (B, arrowhead) are found next to each other and almost never overlap. (C) The Delta protein appears as punctate cytoplasmic staining. The β-galactosidase staining is both cytoplasmic and nuclear, thus overlaps extensively with DAPI staining (blue). In pathogen fed flies, the β-galactosidase staining increased substantially, consistent with the accumulation of more enteroblasts surrounding Delta-positive ISCs. There was also more obvious β-galactosidase staining (red) in cytoplasm, suggesting the cell size of enteroblasts has also increased. However, all Delta-positive cells clearly had no cytoplasmic β-galactosidase staining (indicated by arrows in parts D–O), and appeared to have empty space surrounding the nuclei. Over 100 Delta positive cells were counted in each experiment and no overlap of the staining was observed.
    Figure Legend Snippet: Pathogen feeding does not alter the cell fate decision. Dissected guts from Su(H)-lacZ flies fed with the various agents as indicated were used for immunofluorescent staining. Delta staining (green) and β-galactosidase staining (red) were performed together on the guts. Representative confocal images are shown here. In control samples, the Delta-positive cells (A, arrow) and the Su(H)-lacZ positive cells (B, arrowhead) are found next to each other and almost never overlap. (C) The Delta protein appears as punctate cytoplasmic staining. The β-galactosidase staining is both cytoplasmic and nuclear, thus overlaps extensively with DAPI staining (blue). In pathogen fed flies, the β-galactosidase staining increased substantially, consistent with the accumulation of more enteroblasts surrounding Delta-positive ISCs. There was also more obvious β-galactosidase staining (red) in cytoplasm, suggesting the cell size of enteroblasts has also increased. However, all Delta-positive cells clearly had no cytoplasmic β-galactosidase staining (indicated by arrows in parts D–O), and appeared to have empty space surrounding the nuclei. Over 100 Delta positive cells were counted in each experiment and no overlap of the staining was observed.

    Techniques Used: Staining

    13) Product Images from "Multiple embryonic origins of nitric oxide synthase-expressing GABAergic neurons of the neocortex"

    Article Title: Multiple embryonic origins of nitric oxide synthase-expressing GABAergic neurons of the neocortex

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00065

    Embryonic origin of type I and type II nNOS cortical interneurons. (A) Coexpression of nNOS and GFP/YFP/Venus/β-gal in type I cells in different transgenic mouse lines. Arrows and arrowheads point to double and single-labeled type I cells, respectively. (B) Contribution of different progenitor pools to type I cortical interneurons. The extent of co-localization between nNOS and GFP/YFP/Venus/β-gal in type I cells is shown as a percentage of the total nNOS type I cells (B) or as a percentage of the total GFP/YFP/Venus/ β-gal-expressing interneurons (C) . (D) Coexpression of nNOS and GFP/YFP/Venus/β-gal in type II cells in different transgenic mouse lines. Arrows and arrowheads point to double and single-labeled type II cells, respectively. (E) Contribution of different progenitor pools to type II cells. The extent of co-localization between nNOS and GFP/YFP/Venus/β-gal in type II cells is shown as a percentage of the total nNOS type II cells ( E ) or as a percentage of the total GFP/YFP/Venus/β-gal-expressing interneurons (F) . Error bars indicate SEM. Scale bar: 20 μm.
    Figure Legend Snippet: Embryonic origin of type I and type II nNOS cortical interneurons. (A) Coexpression of nNOS and GFP/YFP/Venus/β-gal in type I cells in different transgenic mouse lines. Arrows and arrowheads point to double and single-labeled type I cells, respectively. (B) Contribution of different progenitor pools to type I cortical interneurons. The extent of co-localization between nNOS and GFP/YFP/Venus/β-gal in type I cells is shown as a percentage of the total nNOS type I cells (B) or as a percentage of the total GFP/YFP/Venus/ β-gal-expressing interneurons (C) . (D) Coexpression of nNOS and GFP/YFP/Venus/β-gal in type II cells in different transgenic mouse lines. Arrows and arrowheads point to double and single-labeled type II cells, respectively. (E) Contribution of different progenitor pools to type II cells. The extent of co-localization between nNOS and GFP/YFP/Venus/β-gal in type II cells is shown as a percentage of the total nNOS type II cells ( E ) or as a percentage of the total GFP/YFP/Venus/β-gal-expressing interneurons (F) . Error bars indicate SEM. Scale bar: 20 μm.

    Techniques Used: Transgenic Assay, Labeling, Expressing

    Laminar distribution of nNOS immunoreactive cells originating in different embryonic neuroepithelial regions. The density of type I (A–F) and type II cells (G–L) coexpressing GFP/YFP/Venus/β-gal (yellow) as well as the total number of nNOS-expressing cells (gray) across cortical bins are shown for each transgenic mouse line. Error bars indicate SEM.
    Figure Legend Snippet: Laminar distribution of nNOS immunoreactive cells originating in different embryonic neuroepithelial regions. The density of type I (A–F) and type II cells (G–L) coexpressing GFP/YFP/Venus/β-gal (yellow) as well as the total number of nNOS-expressing cells (gray) across cortical bins are shown for each transgenic mouse line. Error bars indicate SEM.

    Techniques Used: Expressing, Transgenic Assay

    14) Product Images from "Retromer Controls Planar Polarity Protein Levels and Asymmetric Localization at Intercellular Junctions"

    Article Title: Retromer Controls Planar Polarity Protein Levels and Asymmetric Localization at Intercellular Junctions

    Journal: Current Biology

    doi: 10.1016/j.cub.2018.12.027

    Snx27 Regulates Junctional Levels of Fmi and Stbm via the PDZ Binding Motif of Fmi (A and D) 28-hr APF (A) or 32-hr APF (D) pupal wings carrying clones of Snx27 ( Figure S2 A), marked by loss of GFP immunolabeling (green in A) or RFP fluorescence (red in D). Wings are immunolabeled for Fmi in red and Stbm in blue (A) or Fmi in blue and phalloidin in green (D). Scale bar 10 μm. (B) Quantitation of mean intensity of Fmi (red dots) or Stbm (orange dots) membrane labeling in pupal wing clones of Snx27 . Intensity is shown as a ratio of signal in Snx27 mutant compared to wild-type in each wing. (C) Mean polarity and variation in polarity angle of wings immunolabeled for Fmi in wild-type and Snx27 mutant tissue. (E) 28-hr APF pupal wing with twin clones of arm-PRO-EGFP-fmi next to arm-PRO-EGFP-fmiΔPDZ binding motif (ΔPDZbm), marked by β-gal immunolabeling in blue, in a fmi E59 /fmi E45 mutant background. The wing is immunolabeled for EGFP in green and Stbm in red. (F) Quantitation of mean intensity of membrane labeling of EGFP-Fmi (red dots) and Stbm (orange dots). Intensity is shown as a ratio of signal in ΔPDZbm compared to full-length protein in each wing. (G) Mean polarity and variation in polarity angle of wings immunolabeled for EGFP in EGFP-fmi and EGFP-fmiΔPDZbm tissue. (H) Quantitation of mean intensity of EGFP-Fmi membrane labeling. Intensity is shown as a ratio of EGFP signal in Snx27 mutant compared to wild-type in each wing. (I and J) 28-hr APF pupal wings expressing Arm-PRO-EGFP-fmi (I) or Arm-PRO-EGFP-fmiΔPDZ binding motif ( ΔPDZbm ) (J) and carrying clones of Snx27 , marked by loss of RFP (red). EGFP immunolabeling is in green. (B, F, and H) Error bars are SD. One-sample t tests were used to determine whether the ratio differed from 1.0; ∗∗∗ p
    Figure Legend Snippet: Snx27 Regulates Junctional Levels of Fmi and Stbm via the PDZ Binding Motif of Fmi (A and D) 28-hr APF (A) or 32-hr APF (D) pupal wings carrying clones of Snx27 ( Figure S2 A), marked by loss of GFP immunolabeling (green in A) or RFP fluorescence (red in D). Wings are immunolabeled for Fmi in red and Stbm in blue (A) or Fmi in blue and phalloidin in green (D). Scale bar 10 μm. (B) Quantitation of mean intensity of Fmi (red dots) or Stbm (orange dots) membrane labeling in pupal wing clones of Snx27 . Intensity is shown as a ratio of signal in Snx27 mutant compared to wild-type in each wing. (C) Mean polarity and variation in polarity angle of wings immunolabeled for Fmi in wild-type and Snx27 mutant tissue. (E) 28-hr APF pupal wing with twin clones of arm-PRO-EGFP-fmi next to arm-PRO-EGFP-fmiΔPDZ binding motif (ΔPDZbm), marked by β-gal immunolabeling in blue, in a fmi E59 /fmi E45 mutant background. The wing is immunolabeled for EGFP in green and Stbm in red. (F) Quantitation of mean intensity of membrane labeling of EGFP-Fmi (red dots) and Stbm (orange dots). Intensity is shown as a ratio of signal in ΔPDZbm compared to full-length protein in each wing. (G) Mean polarity and variation in polarity angle of wings immunolabeled for EGFP in EGFP-fmi and EGFP-fmiΔPDZbm tissue. (H) Quantitation of mean intensity of EGFP-Fmi membrane labeling. Intensity is shown as a ratio of EGFP signal in Snx27 mutant compared to wild-type in each wing. (I and J) 28-hr APF pupal wings expressing Arm-PRO-EGFP-fmi (I) or Arm-PRO-EGFP-fmiΔPDZ binding motif ( ΔPDZbm ) (J) and carrying clones of Snx27 , marked by loss of RFP (red). EGFP immunolabeling is in green. (B, F, and H) Error bars are SD. One-sample t tests were used to determine whether the ratio differed from 1.0; ∗∗∗ p

    Techniques Used: Binding Assay, Clone Assay, Immunolabeling, Fluorescence, Quantitation Assay, Labeling, Mutagenesis, Expressing

    Vps35 Regulates Levels of Fmi and Stbm at Apical Junctions Independently of the Wash Complex (A) Diagram illustrating asymmetric localization of the core planar polarity proteins in the pupal wing. Two cells are shown, with Fmi, Fz, Dsh, and Dgo localizing on distal cell edges. This forms an intercellular complex with Fmi, Stbm, and Pk on proximal edges of the neighboring cell. (B) During polarization, complexes sort from a uniform distribution (left), and all the complexes become oriented in the same direction (right). This specifies positioning of trichomes (black in right diagram) to distal cell edges. (C, F, G, and H) 28-hr after puparium formation (APF) (C, G, and H) or 32-hr APF (F) pupal wings carrying clones of Vps35 (C and F), Fam21 (G), or wash (H), marked by loss of β-gal staining (blue in C, G, and H and green in F). Wings are immunolabeled for Fmi in green and Stbm in red (C, G, and H) or Fmi in blue and phalloidin in red (F). The reduced phalloidin staining in Vps35 mutant tissue (F) indicates a delay in trichome initiation. In older wings, phalloidin-stained trichomes are visible in Vps35 clones (not shown). Scale bar 10 μm. (C’) High magnification image of wild-type and mutant regions immunolabeled with Fmi and used to quantitate polarity. (C’’) Polarity nematic showing the magnitude and angle of polarization for each cell. (D) Quantitation of mean intensity of Fmi (red dots) or Stbm (orange dots) membrane labeling in pupal wing clones. Intensity is shown as a ratio of signal in mutant compared to wild-type in each wing; error bars are SD. One-sample t tests were used to determine whether the ratio differed from 1.0. (E) Mean polarity and variation in polarity angle of wings immunolabeled for Fmi in wild-type and Vps35 mutant tissue (see C’ and C’’). Values from the same wing are linked by black bars; mean and SD are listed. Paired t tests were used to compare values in the same wing. ∗∗∗ p
    Figure Legend Snippet: Vps35 Regulates Levels of Fmi and Stbm at Apical Junctions Independently of the Wash Complex (A) Diagram illustrating asymmetric localization of the core planar polarity proteins in the pupal wing. Two cells are shown, with Fmi, Fz, Dsh, and Dgo localizing on distal cell edges. This forms an intercellular complex with Fmi, Stbm, and Pk on proximal edges of the neighboring cell. (B) During polarization, complexes sort from a uniform distribution (left), and all the complexes become oriented in the same direction (right). This specifies positioning of trichomes (black in right diagram) to distal cell edges. (C, F, G, and H) 28-hr after puparium formation (APF) (C, G, and H) or 32-hr APF (F) pupal wings carrying clones of Vps35 (C and F), Fam21 (G), or wash (H), marked by loss of β-gal staining (blue in C, G, and H and green in F). Wings are immunolabeled for Fmi in green and Stbm in red (C, G, and H) or Fmi in blue and phalloidin in red (F). The reduced phalloidin staining in Vps35 mutant tissue (F) indicates a delay in trichome initiation. In older wings, phalloidin-stained trichomes are visible in Vps35 clones (not shown). Scale bar 10 μm. (C’) High magnification image of wild-type and mutant regions immunolabeled with Fmi and used to quantitate polarity. (C’’) Polarity nematic showing the magnitude and angle of polarization for each cell. (D) Quantitation of mean intensity of Fmi (red dots) or Stbm (orange dots) membrane labeling in pupal wing clones. Intensity is shown as a ratio of signal in mutant compared to wild-type in each wing; error bars are SD. One-sample t tests were used to determine whether the ratio differed from 1.0. (E) Mean polarity and variation in polarity angle of wings immunolabeled for Fmi in wild-type and Vps35 mutant tissue (see C’ and C’’). Values from the same wing are linked by black bars; mean and SD are listed. Paired t tests were used to compare values in the same wing. ∗∗∗ p

    Techniques Used: Clone Assay, Staining, Immunolabeling, Mutagenesis, Quantitation Assay, Labeling

    15) Product Images from "Semaphorin 4C Plexin-B2 signaling in peripheral sensory neurons is pronociceptive in a model of inflammatory pain"

    Article Title: Semaphorin 4C Plexin-B2 signaling in peripheral sensory neurons is pronociceptive in a model of inflammatory pain

    Journal: Nature Communications

    doi: 10.1038/s41467-017-00341-w

    Analysis of Plexin-B2 expression in sensory neurons of the dorsal root ganglia (DRG) in adult mice and its regulation in inflammatory pain. a , b Expression of plxnb2 via immunofluorescence analysis of β-galactosidase in adult DRG in respective LacZ reporter knock-in mice. Typical examples a and quantitative analysis b of the distribution of DRG cell types expressing plxnb2 via co-immunolabeling with marker proteins ( n = 10–20 sections/group taken from at least three different mice/group). Scale bar , 50 µm. c , d Typical examples c and quantitative summary d of LacZ staining demonstrating plxnb2 expression in adult Plexin-B2-LacZ +/− mice following intraplantar injection of either vehicle (control) or Complete Freund´s Adjuvant (CFA) stimulating inflammatory pain ( n = 20–30 sections/group taken from at least 3 different mice/group). Scale bars represent 50 µm. In d , Student’s t -test (two sides) was performed. P
    Figure Legend Snippet: Analysis of Plexin-B2 expression in sensory neurons of the dorsal root ganglia (DRG) in adult mice and its regulation in inflammatory pain. a , b Expression of plxnb2 via immunofluorescence analysis of β-galactosidase in adult DRG in respective LacZ reporter knock-in mice. Typical examples a and quantitative analysis b of the distribution of DRG cell types expressing plxnb2 via co-immunolabeling with marker proteins ( n = 10–20 sections/group taken from at least three different mice/group). Scale bar , 50 µm. c , d Typical examples c and quantitative summary d of LacZ staining demonstrating plxnb2 expression in adult Plexin-B2-LacZ +/− mice following intraplantar injection of either vehicle (control) or Complete Freund´s Adjuvant (CFA) stimulating inflammatory pain ( n = 20–30 sections/group taken from at least 3 different mice/group). Scale bars represent 50 µm. In d , Student’s t -test (two sides) was performed. P

    Techniques Used: Expressing, Mouse Assay, Immunofluorescence, Knock-In, Immunolabeling, Marker, Staining, Injection

    Sema4C is expressed in adult DRG and paw skin and plays a functional role in inflammatory pain. a Expression of Sema4c via β-galactosidase (LacZ) staining in adult DRG and plantar paw skin using LacZ reporter knock-in mice. Typical examples ( upper ) and quantitative summary (panel) of LacZ staining in the basal state or following CFA-induced paw inflammation. Scale bars , 50 µm. b Expression of Sema4C and its co-localization with immune cells markers via double immunofluorescence staining in plantar paw skin of mice at 24 h after vehicle or CFA injection, using anti-Sema4C antibody and antibodies against immune cells (CD3 to mark infiltrating T cells, upper ; or GR-1 to target macrophages, lower ). Higher magnification views of dermis are shown on extreme right to illustrate cells showing co-localization ( arrowheads ). Scale bars , 25 µm. c Quantitative measurement of intensity of Sema4C immunoreactivity paw tissue 24 h after intraplantar injection of vehicle or CFA; n = 3 mice/group. d , e Examples ( left ) and densitometric quantifications ( right ) of western blot analysis of Sema4C signal in lysates of L3-L4 DRGs d or paw tissue e 24 h after intraplantar CFA injection; n = 8 for DRGs, n = 9 for paw tissues. f Analysis of inflammatory mechanical hypersensitivity following hindpaw CFA injection in mice lacking Sema4C (Sema4C −/− ) and their wild-type littermates. Frequency of paw withdrawal in response to application of 0.07 g force via a von Frey filament is shown. g Changes in paw response latency to radiant heat following CFA injection in Sema4C −/− mice and their wild-type controls littermates. n = 5 (Sema4C −/− mice) and n = 7 (wild-type littermates) mice/group. Student’s t -test was performed in a – e and two-way ANOVA for repeated measures followed by Tukey’s test was performed in f and g . In f , P
    Figure Legend Snippet: Sema4C is expressed in adult DRG and paw skin and plays a functional role in inflammatory pain. a Expression of Sema4c via β-galactosidase (LacZ) staining in adult DRG and plantar paw skin using LacZ reporter knock-in mice. Typical examples ( upper ) and quantitative summary (panel) of LacZ staining in the basal state or following CFA-induced paw inflammation. Scale bars , 50 µm. b Expression of Sema4C and its co-localization with immune cells markers via double immunofluorescence staining in plantar paw skin of mice at 24 h after vehicle or CFA injection, using anti-Sema4C antibody and antibodies against immune cells (CD3 to mark infiltrating T cells, upper ; or GR-1 to target macrophages, lower ). Higher magnification views of dermis are shown on extreme right to illustrate cells showing co-localization ( arrowheads ). Scale bars , 25 µm. c Quantitative measurement of intensity of Sema4C immunoreactivity paw tissue 24 h after intraplantar injection of vehicle or CFA; n = 3 mice/group. d , e Examples ( left ) and densitometric quantifications ( right ) of western blot analysis of Sema4C signal in lysates of L3-L4 DRGs d or paw tissue e 24 h after intraplantar CFA injection; n = 8 for DRGs, n = 9 for paw tissues. f Analysis of inflammatory mechanical hypersensitivity following hindpaw CFA injection in mice lacking Sema4C (Sema4C −/− ) and their wild-type littermates. Frequency of paw withdrawal in response to application of 0.07 g force via a von Frey filament is shown. g Changes in paw response latency to radiant heat following CFA injection in Sema4C −/− mice and their wild-type controls littermates. n = 5 (Sema4C −/− mice) and n = 7 (wild-type littermates) mice/group. Student’s t -test was performed in a – e and two-way ANOVA for repeated measures followed by Tukey’s test was performed in f and g . In f , P

    Techniques Used: Functional Assay, Expressing, Staining, Knock-In, Mouse Assay, Double Immunofluorescence Staining, Injection, Western Blot

    16) Product Images from "A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse"

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    Journal: Biology Open

    doi: 10.1242/bio.026799

    Comparative Dll4 expression in postnatal and adult retinas. (A1-B3) β-gal activity in P1 postnatal retinas from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. Both lines are clearly active in non-endothelial cell types within the retina at this stage. (C1-D3) β-gal activity in P5 postnatal retinas from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. Dll4-BAC-nlacZ is more active than the endogenous reporter in the postnatal retinal arterial endothelium. (E1-F3) β-gal activity is detected in the arterial and capillary endothelium in both (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ retinas at P7. (G1-H3) Dll4 lacZ/+ is enriched in arteries and capillary vessels (and weakly detected in veins), while Dll4-BAC-nlacZ has no activity within the veins in the adult retina. (I1-I6) IHC and indirect immunofluorescent detection of isolectin (I1), smooth muscle actin (I2), β-gal (I3), and merged (I4-I6) images from representative Dll4-BAC-nlacZ adult retinas. a, artery; on, optic nerve; v, vein. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression in postnatal and adult retinas. (A1-B3) β-gal activity in P1 postnatal retinas from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. Both lines are clearly active in non-endothelial cell types within the retina at this stage. (C1-D3) β-gal activity in P5 postnatal retinas from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. Dll4-BAC-nlacZ is more active than the endogenous reporter in the postnatal retinal arterial endothelium. (E1-F3) β-gal activity is detected in the arterial and capillary endothelium in both (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ retinas at P7. (G1-H3) Dll4 lacZ/+ is enriched in arteries and capillary vessels (and weakly detected in veins), while Dll4-BAC-nlacZ has no activity within the veins in the adult retina. (I1-I6) IHC and indirect immunofluorescent detection of isolectin (I1), smooth muscle actin (I2), β-gal (I3), and merged (I4-I6) images from representative Dll4-BAC-nlacZ adult retinas. a, artery; on, optic nerve; v, vein. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Mouse Assay, Immunohistochemistry

    Comparative Dll4 expression during intermediate and late-stage embryonic development. (A-C) lacZ activity in E12.5 (A), E14.5 (B), and E18.5 (C) Dll4 lacZ/+ mouse embryos and yolk sacs. (D) Intra-littermate body measurements in a Dll4 lacZ/+ litter. Data are presented as averages ±s.e.m. Comparisons were made by Student's t -test (** P =0.0058). (E-G″) lacZ activity in E12.5, E14.5, and E18.5 Dll4-BAC-nlacZ mouse embryos (E-G) and yolk sacs (E′-G″). (H) Intra-littermate body measurements in a Dll4-BAC-nlacZ litter. Data are presented as averages ±s.e.m.; ns, nonsignificant. Comparisons were made by Student's t -test. Noticeable size differences can be observed between genotypes due to heterozygous Dll4 loss of function. (I-J) β-gal IHC on E14.5 skin from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ embryos. I′ and J′ are magnified views of a respective region shown in corresponding panels I and J. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression during intermediate and late-stage embryonic development. (A-C) lacZ activity in E12.5 (A), E14.5 (B), and E18.5 (C) Dll4 lacZ/+ mouse embryos and yolk sacs. (D) Intra-littermate body measurements in a Dll4 lacZ/+ litter. Data are presented as averages ±s.e.m. Comparisons were made by Student's t -test (** P =0.0058). (E-G″) lacZ activity in E12.5, E14.5, and E18.5 Dll4-BAC-nlacZ mouse embryos (E-G) and yolk sacs (E′-G″). (H) Intra-littermate body measurements in a Dll4-BAC-nlacZ litter. Data are presented as averages ±s.e.m.; ns, nonsignificant. Comparisons were made by Student's t -test. Noticeable size differences can be observed between genotypes due to heterozygous Dll4 loss of function. (I-J) β-gal IHC on E14.5 skin from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ embryos. I′ and J′ are magnified views of a respective region shown in corresponding panels I and J. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Immunohistochemistry

    β-gal reporter activity is restricted to arterial vasculature in the skin. (A1-A6) Single channel views of indirect immunofluorescence for CD31 (A1), Dll4 (A2), β-gal (A3), and merged (A4-A6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4 lacZ/+ mouse skin. (B1-B6) CD31 (B1), Podoplanin (B2), β-gal (B3), and merged (B4-B6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4 lacZ/+ mouse skin. (C1-C6) CD31 (C1), SMA (C2), β-gal (C3), and merged (C4-C6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker, smooth muscle actin (SMA). (D1-D6) CD31 (D1), Tuj1 (D2), β-gal (D3), and merged (D4-D6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. (E1-E6) CD31 (E1), Dll4 (E2), β-gal (E3), and merged (E4-E6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4-BAC-nlacZ mouse skin. (F1-F6) CD31 (F1), Podoplanin (F2), β-gal (F3), and merged (F4-F6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4-BAC-nlacZ mouse skin. (G1-G6) CD31 (G1), SMA (G2), β-gal (G3), and merged (G4-G6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker SMA. (H1-H6) CD31 (H1), Tuj1 (H2), β-gal (H3), and merged (H4-H6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. Units depicted are in μm.
    Figure Legend Snippet: β-gal reporter activity is restricted to arterial vasculature in the skin. (A1-A6) Single channel views of indirect immunofluorescence for CD31 (A1), Dll4 (A2), β-gal (A3), and merged (A4-A6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4 lacZ/+ mouse skin. (B1-B6) CD31 (B1), Podoplanin (B2), β-gal (B3), and merged (B4-B6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4 lacZ/+ mouse skin. (C1-C6) CD31 (C1), SMA (C2), β-gal (C3), and merged (C4-C6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker, smooth muscle actin (SMA). (D1-D6) CD31 (D1), Tuj1 (D2), β-gal (D3), and merged (D4-D6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. (E1-E6) CD31 (E1), Dll4 (E2), β-gal (E3), and merged (E4-E6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4-BAC-nlacZ mouse skin. (F1-F6) CD31 (F1), Podoplanin (F2), β-gal (F3), and merged (F4-F6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4-BAC-nlacZ mouse skin. (G1-G6) CD31 (G1), SMA (G2), β-gal (G3), and merged (G4-G6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker SMA. (H1-H6) CD31 (H1), Tuj1 (H2), β-gal (H3), and merged (H4-H6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. Units depicted are in μm.

    Techniques Used: Activity Assay, Immunofluorescence, Marker, BAC Assay

    Comparative Dll4 expression in intermediate and late-stage embryonic hearts and lungs. (A1-B5) β-gal activity in E12.5 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5. β-gal activity is present within coronary plexus (A1,A2,B1,B2), the endocardium of the distal end (A1,B1) and root of the aorta (A4,B4) and pulmonary artery in both lines, as well as the endocardium and subepicardial vasculature (A5,B5), but absent from the epicardium and myocardium. (C1-D5) β-gal activity in E14.5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5. β-gal activity is localized to the endocardium of the aorta in both lines (C4,D4), as well as the chamber endocardium (C5,D5), and subepicardial coronary vasculature (C5,D5). β-gal activity was also detected within a small fraction of the myocardium in the BAC reporter line at this stage. (E1-F5) β-gal activity in E18.5 hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root in Dll4 lacZ/+ animals, but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5), and sparsely in the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle. (G1-H4) β-gal activity in E12.5 lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 are representative cross-sections through the lungs, and boxed in areas are magnified in G4 and H4, revealing activity within the endothelium. (I1-J4) β-gal activity in E14.5 lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-J2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4, revealing endothelial-specific activity in both lines. (K1-L4) β-gal activity in E18.5 lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4, demonstrating endothelial-specific activity in both lines. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression in intermediate and late-stage embryonic hearts and lungs. (A1-B5) β-gal activity in E12.5 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5. β-gal activity is present within coronary plexus (A1,A2,B1,B2), the endocardium of the distal end (A1,B1) and root of the aorta (A4,B4) and pulmonary artery in both lines, as well as the endocardium and subepicardial vasculature (A5,B5), but absent from the epicardium and myocardium. (C1-D5) β-gal activity in E14.5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5. β-gal activity is localized to the endocardium of the aorta in both lines (C4,D4), as well as the chamber endocardium (C5,D5), and subepicardial coronary vasculature (C5,D5). β-gal activity was also detected within a small fraction of the myocardium in the BAC reporter line at this stage. (E1-F5) β-gal activity in E18.5 hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root in Dll4 lacZ/+ animals, but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5), and sparsely in the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle. (G1-H4) β-gal activity in E12.5 lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 are representative cross-sections through the lungs, and boxed in areas are magnified in G4 and H4, revealing activity within the endothelium. (I1-J4) β-gal activity in E14.5 lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-J2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4, revealing endothelial-specific activity in both lines. (K1-L4) β-gal activity in E18.5 lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4, demonstrating endothelial-specific activity in both lines. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression in intermediate and late-stage wholemount embryonic brains. (A1-B9) β-gal activity in the E12.5 embryonic brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the E14.5 embryonic brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the E18.5 embryonic brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; AIC, anterior inferior cerebellar artery; azACA, azygos of the anterior cerebral artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression in intermediate and late-stage wholemount embryonic brains. (A1-B9) β-gal activity in the E12.5 embryonic brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the E14.5 embryonic brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the E18.5 embryonic brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; AIC, anterior inferior cerebellar artery; azACA, azygos of the anterior cerebral artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression during early embryonic development. (A) Schematic of the BAC transgene used for generating the Dll4-BAC-nlacZ mouse line, with a magnified schematic of the nuclear LacZ insertion at the ATG start site of Dll4 . (B-E) β-gal activity in E7.75-E10.5 Dll4 lacZ/+ mouse embryos, ventral (B,C) and sagittal (D,E) views. (F-I) β-gal activity in E7.75-E10.5 Dll4-BAC-nlacZ mouse embryos, ventral (F,G) and sagittal (H,I) views. E′ and I′ show β-gal activity in the embryonic yolk sac. E″ and I″ are magnified views of a representative region shown in corresponding panels E′ and I′, respectively. (J-M) Coronal view of X-gal-stained and Eosin-counterstained sections of E9.5 and E10.5 Dll4 lacZ/+ and Dll4-BAC-nlacZ embryos. aPCs, aortic progenitor cells; CC, cardiac crescent; CV, cardinal vein; DA, dorsal aorta; EC, endocardium; HB, hindbrain; ICA, internal carotid artery; ISA, intersegmental artery; LA, left atrium; LV, left ventricle; NT, neural tube; OFT, outflow tract; PNVP, perineural vascular plexus; RA, right atrium; RV, right ventricle; SV, sinus venosus; black caret, ventral V2 interneuron population. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression during early embryonic development. (A) Schematic of the BAC transgene used for generating the Dll4-BAC-nlacZ mouse line, with a magnified schematic of the nuclear LacZ insertion at the ATG start site of Dll4 . (B-E) β-gal activity in E7.75-E10.5 Dll4 lacZ/+ mouse embryos, ventral (B,C) and sagittal (D,E) views. (F-I) β-gal activity in E7.75-E10.5 Dll4-BAC-nlacZ mouse embryos, ventral (F,G) and sagittal (H,I) views. E′ and I′ show β-gal activity in the embryonic yolk sac. E″ and I″ are magnified views of a representative region shown in corresponding panels E′ and I′, respectively. (J-M) Coronal view of X-gal-stained and Eosin-counterstained sections of E9.5 and E10.5 Dll4 lacZ/+ and Dll4-BAC-nlacZ embryos. aPCs, aortic progenitor cells; CC, cardiac crescent; CV, cardinal vein; DA, dorsal aorta; EC, endocardium; HB, hindbrain; ICA, internal carotid artery; ISA, intersegmental artery; LA, left atrium; LV, left ventricle; NT, neural tube; OFT, outflow tract; PNVP, perineural vascular plexus; RA, right atrium; RV, right ventricle; SV, sinus venosus; black caret, ventral V2 interneuron population. Units depicted are in μm.

    Techniques Used: Expressing, BAC Assay, Activity Assay, Staining

    Comparative Dll4 expression in wholemount postnatal and adult brains. (A1-B3) β-gal activity in the P1 postnatal brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the P5 postnatal brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the adult brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; azACA, azygos of the anterior cerebral artery; AIC, anterior inferior cerebellar artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression in wholemount postnatal and adult brains. (A1-B3) β-gal activity in the P1 postnatal brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the P5 postnatal brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the adult brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; azACA, azygos of the anterior cerebral artery; AIC, anterior inferior cerebellar artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression in postnatal and adult hearts and lungs. (A1-B5) β-gal activity in P1 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5, with activity evident within the endocardial lining of the aorta in both lines (A4,B4), as well as the endocardium, coronary vasculature, and myocardium (A5,B5). (C1-D5) β-gal activity in P5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5, with signal evident within the endocardial lining of the aorta in both lines, and persisting in the endocardium of the aorta (C4,D4) and chambers, as well as the myocardium and coronary vasculature (C5,D5). (E1-F5) β-gal activity in adult hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from adult Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, magnified in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root Dll4 lacZ/+ but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5) (asterisks), and within the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle; sm, smooth muscle; asterisks – denote lumenized vasculature. (G1-H4) β-gal activity in P1 postnatal lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels G4 and H4. (I1-J4) β-gal activity in P5 postnatal lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-D2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4. (K1-L4) β-gal activity in adult lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4. In both lines, and at all stages, β-gal activity appears to be confined to the endothelium. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.
    Figure Legend Snippet: Comparative Dll4 expression in postnatal and adult hearts and lungs. (A1-B5) β-gal activity in P1 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5, with activity evident within the endocardial lining of the aorta in both lines (A4,B4), as well as the endocardium, coronary vasculature, and myocardium (A5,B5). (C1-D5) β-gal activity in P5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5, with signal evident within the endocardial lining of the aorta in both lines, and persisting in the endocardium of the aorta (C4,D4) and chambers, as well as the myocardium and coronary vasculature (C5,D5). (E1-F5) β-gal activity in adult hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from adult Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, magnified in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root Dll4 lacZ/+ but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5) (asterisks), and within the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle; sm, smooth muscle; asterisks – denote lumenized vasculature. (G1-H4) β-gal activity in P1 postnatal lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels G4 and H4. (I1-J4) β-gal activity in P5 postnatal lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-D2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4. (K1-L4) β-gal activity in adult lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4. In both lines, and at all stages, β-gal activity appears to be confined to the endothelium. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Techniques Used: Expressing, Activity Assay, BAC Assay, Mouse Assay

    17) Product Images from "The ALS8 protein VAPB interacts with the ER\u2013Golgi recycling protein YIF1A and regulates membrane delivery into dendrites"

    Article Title: The ALS8 protein VAPB interacts with the ER\u2013Golgi recycling protein YIF1A and regulates membrane delivery into dendrites

    Journal: The EMBO Journal

    doi: 10.1038/emboj.2013.131

    YIF1 and VAP are required for normal dendrite morphology. ( A ) Hippocampal neurons co-transfected at DIV1 with indicated constructs and β-galactosidase to visualize morphology. ( B ) Representative image of a hippocampal neurons (DIV5) co-transfected
    Figure Legend Snippet: YIF1 and VAP are required for normal dendrite morphology. ( A ) Hippocampal neurons co-transfected at DIV1 with indicated constructs and β-galactosidase to visualize morphology. ( B ) Representative image of a hippocampal neurons (DIV5) co-transfected

    Techniques Used: Transfection, Construct

    18) Product Images from "Epithelial cell-turnover ensures robust coordination of tissue growth in Drosophila ribosomal protein mutants"

    Article Title: Epithelial cell-turnover ensures robust coordination of tissue growth in Drosophila ribosomal protein mutants

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1009300

    Aberrant Wg signaling gradient is essential for the induction of massive cell-turnover. (A-B”) The relative position between dying cells (cleaved-PARP, magenta) and Wg expression (anti-Wg, green) in the wild-type (A) or RpS3/+ (B) wing pouch. Scale bar, 100 μm. (C-D”) The relative position between dying cells (cleaved-PARP, magenta) and Wg signaling activity (anti-β-galactosidase, green) in the nmo-LacZ /+ (C) or RpS3/+ , nmo-LacZ /+ (D) wing pouch. These images are Z-stacked images from 8 (C) and 8 (D) confocal images (3.02 μm and 2.39 μm, respectively) for whole wing pouch that is acquired every 1.01 μm or 0.8 μm, respectively. Scale bar, 50 μm. (E-F’) RpS3 was overexpressed in the posterior compartment of wing discs of nmo-lacZ/+ (E) or RpS3/+ , nmo-lacZ/+ (F) flies using the en-Gal4 driver. nmo expression was visualized by anti-β-galactosidase staining (white). Scale bar, 100 μm. (G-H’) RpS3 was overexpressed in the posterior compartment of wing discs of nkd-lacZ/+ (G) or RpS3/+ , nkd-lacZ/+ (H) flies using the en-Gal4 driver. nkd expression was visualized by anti-β-galactosidase staining (white). Scale bar, 100 μm. (I-K’) Dying cells were visualized by anti-cleaved PARP staining (white) in the wing discs of RpS3/+ , wg /+ (I), RpS3/+ , nub-Gal4 , UAS-Wg (J), or RpS3/+ , nub-Gal4 , UAS-dsh-RNAi (K) flies expressing CD8-PARP-Venus. Wg expression was visualized by anti-Wg staining (white). Scale bar, 100 μm. (L) Boxplot with dots representing cleaved-PARP-positive dying cells per pouch in genotypes shown in (B) (n = 12, number of wing pouches), (I) (n = 9), (J) (n = 10), and (K) (n = 11). Error bars, SEM; ***, p
    Figure Legend Snippet: Aberrant Wg signaling gradient is essential for the induction of massive cell-turnover. (A-B”) The relative position between dying cells (cleaved-PARP, magenta) and Wg expression (anti-Wg, green) in the wild-type (A) or RpS3/+ (B) wing pouch. Scale bar, 100 μm. (C-D”) The relative position between dying cells (cleaved-PARP, magenta) and Wg signaling activity (anti-β-galactosidase, green) in the nmo-LacZ /+ (C) or RpS3/+ , nmo-LacZ /+ (D) wing pouch. These images are Z-stacked images from 8 (C) and 8 (D) confocal images (3.02 μm and 2.39 μm, respectively) for whole wing pouch that is acquired every 1.01 μm or 0.8 μm, respectively. Scale bar, 50 μm. (E-F’) RpS3 was overexpressed in the posterior compartment of wing discs of nmo-lacZ/+ (E) or RpS3/+ , nmo-lacZ/+ (F) flies using the en-Gal4 driver. nmo expression was visualized by anti-β-galactosidase staining (white). Scale bar, 100 μm. (G-H’) RpS3 was overexpressed in the posterior compartment of wing discs of nkd-lacZ/+ (G) or RpS3/+ , nkd-lacZ/+ (H) flies using the en-Gal4 driver. nkd expression was visualized by anti-β-galactosidase staining (white). Scale bar, 100 μm. (I-K’) Dying cells were visualized by anti-cleaved PARP staining (white) in the wing discs of RpS3/+ , wg /+ (I), RpS3/+ , nub-Gal4 , UAS-Wg (J), or RpS3/+ , nub-Gal4 , UAS-dsh-RNAi (K) flies expressing CD8-PARP-Venus. Wg expression was visualized by anti-Wg staining (white). Scale bar, 100 μm. (L) Boxplot with dots representing cleaved-PARP-positive dying cells per pouch in genotypes shown in (B) (n = 12, number of wing pouches), (I) (n = 9), (J) (n = 10), and (K) (n = 11). Error bars, SEM; ***, p

    Techniques Used: Expressing, Activity Assay, Staining

    19) Product Images from "The role of integrins in Drosophila egg chamber morphogenesis"

    Article Title: The role of integrins in Drosophila egg chamber morphogenesis

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.182774

    Mys is required for interfollicular stalk formation. (A) A wild-type ovariole expressing Mys-GFP (green) from a genomic BAC transgene. Mys is enriched on the basal side of all pre-follicle cells in the germarium (bracket) and in the forming and mature interfollicular stalks (yellow arrowheads and white arrowhead, respectively) ( n =4). (B) A section of ovariole stained for Lgl (red), DNA (blue) and β-galactosidase (green and B′) showing the pattern of UAS-LacZ expression under the control of the 24B-Gal4 driver. 24B-Gal4 drives expression in the interfollicular stalk cells only (arrowhead and arrows) ( n =6). (C) A stage 4 egg chamber stained for Dlg (white) and DNA (blue) containing mys XG43 clones marked by the loss of RFP (red). 24B-Gal4 was used to drive expression of UAS-FLP to generate mys XG43 clones specifically in the interfollicular stalks. The presence of mys mutant cells disrupts the organisation of the interfollicular stalks ( n =14/15). (D) A wild-type stage 4 egg chamber stained for F-actin (white) and DNA (blue) showing the fully formed stalks that separate it from the adjacent younger and older egg chambers ( n =5). (E-J′) For these experiments, mys XG43 clones [marked by the absence of RFP (red)] were induced in interfollicular stalks by the expression UAS-FLP under the control of 24B-Gal4. (E,E′) A disorganised stalk containing mys clones stained for F-actin (green), DNA (blue) and Lamin C (white in E′). Lamin C is expressed in both mutant and wild-type cells in disrupted interfollicular stalks (E) ( n =8). (F,F′) A disorganised stalk containing mys clones stained for Eya (green) and DNA (blue). Eya is turned off normally in both the mutant and wild-type stalk cells ( n =7). (G-I′) Mutant interfollicular stalk cells are round in appearance ( n =33) compared with the stalk made from all wild-type cells (I) ( n =4). Wild-type cells in disrupted interfollicular stalks range from rounded (white asterisks in G,G′) ( n =18/51) to wild type (white asterisks in H,H′) ( n =33/51). Dlg only localises to the regions where neighbouring interfollicular stalk cells contact each other in disrupted interfollicular stalks (arrowheads G′ and H′) ( n =18/18) as in interfollicular stalks containing only wild-type cells (I′) ( n =8/8). (J) A disorganised interfollicular stalk stained for aPKC (green) and Dlg (white). (J′) A z -projection of 16 planes taken 1 μm apart of the stalk in J. This shows that aPKC is not present in mature stalks containing mys XG43 mutant cells ( n =11). (K-L′) Regions of ovarioles stained for DNA (blue) and β-galactosidase (green) containing hs-FLP-induced mys XG43 clones marked by the loss of RFP (red). The interfollicular stalk cells are marked by β-galactosidase (green) expressed from UAS-LacZ under the control of 24B-Gal4. The disorganised region caused by mys XG43 clones at the terminus of the stage 9 egg chamber in the boxed area in K (enlarged in K′) does not contain cells expressing the β-galactosidase stalk marker, which lie only in a malformed stalk posteriorly (K′) ( n =4). (L,L′) The stalk cells lie at the posterior of the region between the two younger egg chambers ( lacZ -positive cells, green). The black dashed line identifies cells that could potentially contribute to the disorganised region at the terminus of the egg chamber later in development, like that seen in K ( n =4). Scale bars: 10 μm.
    Figure Legend Snippet: Mys is required for interfollicular stalk formation. (A) A wild-type ovariole expressing Mys-GFP (green) from a genomic BAC transgene. Mys is enriched on the basal side of all pre-follicle cells in the germarium (bracket) and in the forming and mature interfollicular stalks (yellow arrowheads and white arrowhead, respectively) ( n =4). (B) A section of ovariole stained for Lgl (red), DNA (blue) and β-galactosidase (green and B′) showing the pattern of UAS-LacZ expression under the control of the 24B-Gal4 driver. 24B-Gal4 drives expression in the interfollicular stalk cells only (arrowhead and arrows) ( n =6). (C) A stage 4 egg chamber stained for Dlg (white) and DNA (blue) containing mys XG43 clones marked by the loss of RFP (red). 24B-Gal4 was used to drive expression of UAS-FLP to generate mys XG43 clones specifically in the interfollicular stalks. The presence of mys mutant cells disrupts the organisation of the interfollicular stalks ( n =14/15). (D) A wild-type stage 4 egg chamber stained for F-actin (white) and DNA (blue) showing the fully formed stalks that separate it from the adjacent younger and older egg chambers ( n =5). (E-J′) For these experiments, mys XG43 clones [marked by the absence of RFP (red)] were induced in interfollicular stalks by the expression UAS-FLP under the control of 24B-Gal4. (E,E′) A disorganised stalk containing mys clones stained for F-actin (green), DNA (blue) and Lamin C (white in E′). Lamin C is expressed in both mutant and wild-type cells in disrupted interfollicular stalks (E) ( n =8). (F,F′) A disorganised stalk containing mys clones stained for Eya (green) and DNA (blue). Eya is turned off normally in both the mutant and wild-type stalk cells ( n =7). (G-I′) Mutant interfollicular stalk cells are round in appearance ( n =33) compared with the stalk made from all wild-type cells (I) ( n =4). Wild-type cells in disrupted interfollicular stalks range from rounded (white asterisks in G,G′) ( n =18/51) to wild type (white asterisks in H,H′) ( n =33/51). Dlg only localises to the regions where neighbouring interfollicular stalk cells contact each other in disrupted interfollicular stalks (arrowheads G′ and H′) ( n =18/18) as in interfollicular stalks containing only wild-type cells (I′) ( n =8/8). (J) A disorganised interfollicular stalk stained for aPKC (green) and Dlg (white). (J′) A z -projection of 16 planes taken 1 μm apart of the stalk in J. This shows that aPKC is not present in mature stalks containing mys XG43 mutant cells ( n =11). (K-L′) Regions of ovarioles stained for DNA (blue) and β-galactosidase (green) containing hs-FLP-induced mys XG43 clones marked by the loss of RFP (red). The interfollicular stalk cells are marked by β-galactosidase (green) expressed from UAS-LacZ under the control of 24B-Gal4. The disorganised region caused by mys XG43 clones at the terminus of the stage 9 egg chamber in the boxed area in K (enlarged in K′) does not contain cells expressing the β-galactosidase stalk marker, which lie only in a malformed stalk posteriorly (K′) ( n =4). (L,L′) The stalk cells lie at the posterior of the region between the two younger egg chambers ( lacZ -positive cells, green). The black dashed line identifies cells that could potentially contribute to the disorganised region at the terminus of the egg chamber later in development, like that seen in K ( n =4). Scale bars: 10 μm.

    Techniques Used: Expressing, BAC Assay, Staining, Clone Assay, Mutagenesis, Marker

    20) Product Images from "Viral caspase inhibitor p35, but not crmA, is neuroprotective in the ischemic penumbra following experimental stroke"

    Article Title: Viral caspase inhibitor p35, but not crmA, is neuroprotective in the ischemic penumbra following experimental stroke

    Journal:

    doi: 10.1016/j.neuroscience.2007.07.030

    (A) Double immunofluorescence staining of β-gal and activated caspase 3 from brains survived 48 hr after stroke. Activated caspase-3 was not detected in the non-ischemic cortex (data not shown). Transfected neurons with control, p35 or crmA vectors
    Figure Legend Snippet: (A) Double immunofluorescence staining of β-gal and activated caspase 3 from brains survived 48 hr after stroke. Activated caspase-3 was not detected in the non-ischemic cortex (data not shown). Transfected neurons with control, p35 or crmA vectors

    Techniques Used: Double Immunofluorescence Staining, Transfection

    (A) Double immunofluorescence staining of β-gal and cytochrome c in ischemic brains 48 hr after stroke. Double-stained neurons were marked with arrows and neurons stained with β-gal only were marked with astericks. An inset was inserted
    Figure Legend Snippet: (A) Double immunofluorescence staining of β-gal and cytochrome c in ischemic brains 48 hr after stroke. Double-stained neurons were marked with arrows and neurons stained with β-gal only were marked with astericks. An inset was inserted

    Techniques Used: Double Immunofluorescence Staining, Staining

    (A) Double immunofluorescence staining of β-gal and AIF in ischemic cortex 48 hr after stroke. Positive AIF staining was detected in the ischemic but not non-ischemic cortex (data not shown). After 48 hr of MCAO, most of AIF expression localized
    Figure Legend Snippet: (A) Double immunofluorescence staining of β-gal and AIF in ischemic cortex 48 hr after stroke. Positive AIF staining was detected in the ischemic but not non-ischemic cortex (data not shown). After 48 hr of MCAO, most of AIF expression localized

    Techniques Used: Double Immunofluorescence Staining, Staining, Expressing

    21) Product Images from "Dual Roles for Membrane Association of Drosophila Axin in Wnt Signaling"

    Article Title: Dual Roles for Membrane Association of Drosophila Axin in Wnt Signaling

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1006494

    Membrane-associated Axin is sufficient for Wingless signaling (A-C) Confocal images of third instar larval wing discs with Axin s044230 null mutant clones (marked by the absence of β-gal in A (magenta)). The Wingless target gene senseless (green) is ectopically activated in Axin mutant clones (B and C). (D-F) Expressing Myr-Axin-V5 in wing discs using the 71B-Gal4 driver restores normal senseless expression in Axin null mutant clones. Yellow arrows indicate the dorsoventral boundary. (G-I) Arm is ectopically stabilized in Axin null mutant clones (marked by the absence of β-gal in G). (J-L) Expressing Myr-Axin-V5 in wing discs using the 71B-Gal4 driver suppresses aberrant Arm stabilization in Axin null mutant clones. Scale bar: 20μm.
    Figure Legend Snippet: Membrane-associated Axin is sufficient for Wingless signaling (A-C) Confocal images of third instar larval wing discs with Axin s044230 null mutant clones (marked by the absence of β-gal in A (magenta)). The Wingless target gene senseless (green) is ectopically activated in Axin mutant clones (B and C). (D-F) Expressing Myr-Axin-V5 in wing discs using the 71B-Gal4 driver restores normal senseless expression in Axin null mutant clones. Yellow arrows indicate the dorsoventral boundary. (G-I) Arm is ectopically stabilized in Axin null mutant clones (marked by the absence of β-gal in G). (J-L) Expressing Myr-Axin-V5 in wing discs using the 71B-Gal4 driver suppresses aberrant Arm stabilization in Axin null mutant clones. Scale bar: 20μm.

    Techniques Used: Mutagenesis, Clone Assay, Expressing

    Apc is required for the localization of Axin to puncta juxtaposed with cell membrane. Images of third instar larval wing imaginal discs stained with indicated antibodies; genotypes at left margin. (A-C) Apc2 antibody revealed specific staining of endogenous Apc2 (green), which is absent in Apc2 19 . 3 null mutant clones (marked by the absence of β-gal (magenta), -/- in A). (D-F) Double staining with Fas III (magenta) and Apc2 antibodies indicated that Apc2 partially overlaps Fas III and is enriched at cell cortex. (G-I) Double staining with Apc2 and Axin (magenta) antibodies reveals that Apc2 is present at some membrane-proximal Axin puncta (white arrow), whereas distinct Apc2 or Axin puncta are also observed (yellow and red arrowheads respectively). (J-N) Wing disc triple labeled with β-gal (J, magenta), Axin (K, green) and Fas III (M, red) antibodies. Merge of Axin and β-gal is shown in (L), and merge of Axin and Fas III is in (N). Apc2 19 . 3 homozygous null mutant clones are marked by the absence of β-gal (-/- in J). Endogenous Axin staining indicates reduced Axin puncta at the basolateral membrane in Apc2 19 . 3 null mutant clones (K, L, N). Fas III localization is not disrupted in Apc2 19 . 3 mutant clones (M). (O-S) Wing discs bearing Apc1 Q8 null mutant clones (marked by the absence of β-gal staining, -/- in O) were stained with β-gal (O), Axin (P) and Fas III (R) antibodies. Merge of Axin and β-gal is shown in (Q), and merge of Axin and Fas III is in (S). Axin puncta are reduced at the basolateral membrane in Apc1 Q8 mutant clones (P, Q, S). Fas III localization is not disrupted in Apc1 Q8 mutant clones (R). Scale bar: 5μm.
    Figure Legend Snippet: Apc is required for the localization of Axin to puncta juxtaposed with cell membrane. Images of third instar larval wing imaginal discs stained with indicated antibodies; genotypes at left margin. (A-C) Apc2 antibody revealed specific staining of endogenous Apc2 (green), which is absent in Apc2 19 . 3 null mutant clones (marked by the absence of β-gal (magenta), -/- in A). (D-F) Double staining with Fas III (magenta) and Apc2 antibodies indicated that Apc2 partially overlaps Fas III and is enriched at cell cortex. (G-I) Double staining with Apc2 and Axin (magenta) antibodies reveals that Apc2 is present at some membrane-proximal Axin puncta (white arrow), whereas distinct Apc2 or Axin puncta are also observed (yellow and red arrowheads respectively). (J-N) Wing disc triple labeled with β-gal (J, magenta), Axin (K, green) and Fas III (M, red) antibodies. Merge of Axin and β-gal is shown in (L), and merge of Axin and Fas III is in (N). Apc2 19 . 3 homozygous null mutant clones are marked by the absence of β-gal (-/- in J). Endogenous Axin staining indicates reduced Axin puncta at the basolateral membrane in Apc2 19 . 3 null mutant clones (K, L, N). Fas III localization is not disrupted in Apc2 19 . 3 mutant clones (M). (O-S) Wing discs bearing Apc1 Q8 null mutant clones (marked by the absence of β-gal staining, -/- in O) were stained with β-gal (O), Axin (P) and Fas III (R) antibodies. Merge of Axin and β-gal is shown in (Q), and merge of Axin and Fas III is in (S). Axin puncta are reduced at the basolateral membrane in Apc1 Q8 mutant clones (P, Q, S). Fas III localization is not disrupted in Apc1 Q8 mutant clones (R). Scale bar: 5μm.

    Techniques Used: Staining, Mutagenesis, Clone Assay, Double Staining, Labeling

    Axin is localized at cell membrane-proximal puncta independently of Wingless pathway activation (A-I) Confocal images of third instar larval wing imaginal discs stained with antibodies indicated at bottom right; genotypes at left margin. (A-C) Wing disc stained with β-gal (A, magenta), Axin (B, green), and Arm (C, blue) antibodies. Axin 18 null mutant clones (marked by the absence of β-gal, -/- in A) demonstrate the specificity of the Axin antibody. Armadillo marks the adherens junctions, which are present at the boundary between the apical and basolateral membrane, and also accumulates in the cytoplasm in Axin mutant clones. At this apical level, Axin staining is diffuse in the cytoplasm of all cells. (D-I) Axin staining at basolateral levels in the wing disc. Axin 18 null mutant clones are marked by the absence of β-gal (D) or by dashed line (G). At this level, Axin antibody reveals specific staining that partially overlaps the basolateral membrane marker Fas III (I). Higher magnification views of the boxed area in (H) reveals endogenous Axin is present in puncta at or near the plasma membrane (G’-I’). Images were taken at the periphery of the wing discs. (J-L) Wild-type pupal wing (~28 hrs after pupa formation) double labeled with α-Dlg (J) and α-Axin (K). Endogenous Axin is also present in puncta proximal to the cell membrane in pupal wing (L). (M) Subcellular fractionation of lysates from S2R+ cells. The total lysates, cytoplasmic, and membrane fractions were analyzed by SDS-PAGE. Immunoblotting with Axin antibody revealed that Axin is present in both the cytoplasmic and membrane fractions. The efficiency of the fractionation was assayed by the presence of Arrow and Tubulin, membrane and cytoplasmic markers, respectively. (N) Quantification of the distribution of endogenous Axin in S2R+ cells. Results were obtained from four independent experiments, with a representative blot shown in (M). Values indicate mean ± SD. (O) Subcellular fractionation of lysates from 0–2.5 hour wild-type embryos. (P) Quantification of the distribution of endogenous Axin in 0–2.5 hour wild-type embryos. Results were obtained from four independent experiments, with a representative blot shown in (O). Values indicate mean ± SD. Scale bar: 5μm.
    Figure Legend Snippet: Axin is localized at cell membrane-proximal puncta independently of Wingless pathway activation (A-I) Confocal images of third instar larval wing imaginal discs stained with antibodies indicated at bottom right; genotypes at left margin. (A-C) Wing disc stained with β-gal (A, magenta), Axin (B, green), and Arm (C, blue) antibodies. Axin 18 null mutant clones (marked by the absence of β-gal, -/- in A) demonstrate the specificity of the Axin antibody. Armadillo marks the adherens junctions, which are present at the boundary between the apical and basolateral membrane, and also accumulates in the cytoplasm in Axin mutant clones. At this apical level, Axin staining is diffuse in the cytoplasm of all cells. (D-I) Axin staining at basolateral levels in the wing disc. Axin 18 null mutant clones are marked by the absence of β-gal (D) or by dashed line (G). At this level, Axin antibody reveals specific staining that partially overlaps the basolateral membrane marker Fas III (I). Higher magnification views of the boxed area in (H) reveals endogenous Axin is present in puncta at or near the plasma membrane (G’-I’). Images were taken at the periphery of the wing discs. (J-L) Wild-type pupal wing (~28 hrs after pupa formation) double labeled with α-Dlg (J) and α-Axin (K). Endogenous Axin is also present in puncta proximal to the cell membrane in pupal wing (L). (M) Subcellular fractionation of lysates from S2R+ cells. The total lysates, cytoplasmic, and membrane fractions were analyzed by SDS-PAGE. Immunoblotting with Axin antibody revealed that Axin is present in both the cytoplasmic and membrane fractions. The efficiency of the fractionation was assayed by the presence of Arrow and Tubulin, membrane and cytoplasmic markers, respectively. (N) Quantification of the distribution of endogenous Axin in S2R+ cells. Results were obtained from four independent experiments, with a representative blot shown in (M). Values indicate mean ± SD. (O) Subcellular fractionation of lysates from 0–2.5 hour wild-type embryos. (P) Quantification of the distribution of endogenous Axin in 0–2.5 hour wild-type embryos. Results were obtained from four independent experiments, with a representative blot shown in (O). Values indicate mean ± SD. Scale bar: 5μm.

    Techniques Used: Activation Assay, Staining, Mutagenesis, Clone Assay, Marker, Labeling, Fractionation, SDS Page

    22) Product Images from "RacGAP50C directs perinuclear ?-tubulin localization to organize the uniform microtubule array required for Drosophila myotube extension"

    Article Title: RacGAP50C directs perinuclear ?-tubulin localization to organize the uniform microtubule array required for Drosophila myotube extension

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.031823

    RacGAP DH15 mutants can form stable muscle attachments. Wild-type (A,C) or RacGAP DH15 mutant (B,D) stage 16 Drosophila embryos. To the left is a schematic of the wild-type muscle pattern (magenta) and tendon cells (green). ( A , B ) Muscle myosin and β-galactosidase
    Figure Legend Snippet: RacGAP DH15 mutants can form stable muscle attachments. Wild-type (A,C) or RacGAP DH15 mutant (B,D) stage 16 Drosophila embryos. To the left is a schematic of the wild-type muscle pattern (magenta) and tendon cells (green). ( A , B ) Muscle myosin and β-galactosidase

    Techniques Used: Mutagenesis

    23) Product Images from "Wnt-Responsive Lgr5+ Globose Basal Cells Function as Multipotent Olfactory Epithelium Progenitor Cells"

    Article Title: Wnt-Responsive Lgr5+ Globose Basal Cells Function as Multipotent Olfactory Epithelium Progenitor Cells

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0240-14.2014

    Sustained Wnt activation delays olfactory sensory neuron maturation. A–C , Confocal images of olfactory epithelium stained with antibodies specific for OMP and β-gal. Compared with wild-type control ( A ), downregulation of Wnt activity by
    Figure Legend Snippet: Sustained Wnt activation delays olfactory sensory neuron maturation. A–C , Confocal images of olfactory epithelium stained with antibodies specific for OMP and β-gal. Compared with wild-type control ( A ), downregulation of Wnt activity by

    Techniques Used: Activation Assay, Staining, Activity Assay

    24) Product Images from "Hippo, TGF-β, and Src-MAPK pathways regulate transcription of the upd3 cytokine in Drosophila enterocytes upon bacterial infection"

    Article Title: Hippo, TGF-β, and Src-MAPK pathways regulate transcription of the upd3 cytokine in Drosophila enterocytes upon bacterial infection

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1007091

    The upd3 gene is regulated by cell-specific, region-specific and infection-responsive enhancers. (A) Schematic of the upd3 gene and the 21 overlapping sequences used to create GFP reporter lines. The upd3 exons are represented by orange blocks and the introns are light blue. Putative enhancer regions have been color coded by their ability to drive GFP expression as follows: Solid Grey–no midgut signal, Dashed Grey–infection induced signal in scattered cells, Green–infection-induced signal throughout the gut, Blue–constant signal throughout the gut, Pink–infection induced signal in a specific midgut region, Purple–constant signal confined to a specific midgut region. (B) Enhancer region M drives an unvarying GFP signal in esg-lacZ expressing cells (ISCs and EBs) in all regions. (C, D) Both the C and I enhancer region sequences drive GFP in an infection-inducible manner, specifically in Myo- positive cells (ECs) throughout the midgut. (E) Enhancer region R drives infection-induced GFP expression in esg -positive cells (ISCs and EBs). (B, C, D, E) Confocal microscopy images taken at 40x magnification with four color channels. DAPI stained nuclei in Blue, GFP in green and antibody stained β-Gal in red. Scale bars are 50μm.
    Figure Legend Snippet: The upd3 gene is regulated by cell-specific, region-specific and infection-responsive enhancers. (A) Schematic of the upd3 gene and the 21 overlapping sequences used to create GFP reporter lines. The upd3 exons are represented by orange blocks and the introns are light blue. Putative enhancer regions have been color coded by their ability to drive GFP expression as follows: Solid Grey–no midgut signal, Dashed Grey–infection induced signal in scattered cells, Green–infection-induced signal throughout the gut, Blue–constant signal throughout the gut, Pink–infection induced signal in a specific midgut region, Purple–constant signal confined to a specific midgut region. (B) Enhancer region M drives an unvarying GFP signal in esg-lacZ expressing cells (ISCs and EBs) in all regions. (C, D) Both the C and I enhancer region sequences drive GFP in an infection-inducible manner, specifically in Myo- positive cells (ECs) throughout the midgut. (E) Enhancer region R drives infection-induced GFP expression in esg -positive cells (ISCs and EBs). (B, C, D, E) Confocal microscopy images taken at 40x magnification with four color channels. DAPI stained nuclei in Blue, GFP in green and antibody stained β-Gal in red. Scale bars are 50μm.

    Techniques Used: Infection, Expressing, Confocal Microscopy, Staining

    25) Product Images from "Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism"

    Article Title: Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.071761

    Dll1 , NICD and Hes1 expression in endoderm. ( A-D ) Optical sections of E7.5 to E10.5 Dll1 lacZ /+ embryos whole-mount stained for β-galactosidase, Cdh1, Pdx1, Neurog3 or Nkx6-1. The arrows in C point to Neurog3 + cells that co-express β-gal.
    Figure Legend Snippet: Dll1 , NICD and Hes1 expression in endoderm. ( A-D ) Optical sections of E7.5 to E10.5 Dll1 lacZ /+ embryos whole-mount stained for β-galactosidase, Cdh1, Pdx1, Neurog3 or Nkx6-1. The arrows in C point to Neurog3 + cells that co-express β-gal.

    Techniques Used: Expressing, Staining

    Ptf1a is required for Dll1 expression in MPCs. ( A-F ′) Image stack projections (A-C) and optical sections (D-E) of whole-mount stained E10.5 embryos of the indicated genotypes, stained for β-galactosidase indicating Dll1 expression, Pdx1
    Figure Legend Snippet: Ptf1a is required for Dll1 expression in MPCs. ( A-F ′) Image stack projections (A-C) and optical sections (D-E) of whole-mount stained E10.5 embryos of the indicated genotypes, stained for β-galactosidase indicating Dll1 expression, Pdx1

    Techniques Used: Expressing, Staining

    26) Product Images from "Prostate-specific Klf6 Inactivation Impairs Anterior Prostate Branching Morphogenesis through Increased Activation of the Shh Pathway *"

    Article Title: Prostate-specific Klf6 Inactivation Impairs Anterior Prostate Branching Morphogenesis through Increased Activation of the Shh Pathway *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.001776

    β-Galactosidase expression in Klf6 f/f Nkx3.1 Cre/+ ;Ptc-lacZ reporter mouse prostates. Immunostaining of prostate tissue sections prepared from P7 Klf6 f/f Nkx3.1 Cre/+ ;Ptc-lacZ ( A and C ) and Klf6 f/f Nkx3.1 +/+ ;Ptc-lacZ ( B and D ) mice with anti-β-galactosidase
    Figure Legend Snippet: β-Galactosidase expression in Klf6 f/f Nkx3.1 Cre/+ ;Ptc-lacZ reporter mouse prostates. Immunostaining of prostate tissue sections prepared from P7 Klf6 f/f Nkx3.1 Cre/+ ;Ptc-lacZ ( A and C ) and Klf6 f/f Nkx3.1 +/+ ;Ptc-lacZ ( B and D ) mice with anti-β-galactosidase

    Techniques Used: Expressing, Immunostaining, Mouse Assay

    27) Product Images from "Drosophila Sidekick is required in developing photoreceptors to enable visual motion detection"

    Article Title: Drosophila Sidekick is required in developing photoreceptors to enable visual motion detection

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.158246

    Lamina neuron placement requires sdk in photoreceptors. (A) Diagram of a coronal section through the third-instar larval brain, showing neuroblasts (nb) and lamina precursor cells (LPCs), which become postmitotic (pLPCs) behind the lamina furrow (LF; arrowhead) and differentiate into lamina neurons (LN) aligned into columns along the axons of R1-R6. Glia are shown in blue. (B-E″) Confocal images of the same view of larval brains for: (B-B″) control ( sdk MB05054 /+ ); (C-C″) sdk MB05054 ; (D-D″) sdk knockdown in the eye with ey 3.5 -FLP, Act > CD2 > GAL4 [Sdk still accumulates in LNs (asterisk)]; and (E-E″) sdk knockdown in the lamina with NP6099-GAL4 . A-E″ are labeled with anti-Sdk (B″,C″,D″,E″, green in B′,C′,D′,E′), anti-HRP to mark photoreceptor axons (red in B,B′,C,C′,D,D′,E,E′), anti-β-galactosidase (β-gal) reflecting dac-lacZ (green in B,C) or anti-Dac (green in D,E) to mark lamina neurons, anti-Repo to mark glia (blue in B,C), or anti-β-gal (blue in D,D′) or anti-GFP (blue in E,E′) to mark the domain of RNAi expression. Lamina neurons are misplaced in sdk mutants and when sdk is knocked down in the eye (empty arrowheads, C,D), but not when it is knocked down in lamina neurons or in glia with repo-GAL4 . Filled arrowheads mark the lamina furrow. (F) The number of LNs per µm in or beyond the lamina plexus (LP; arrows in A-E′) in the indicated genotypes. Data are mean±s.e.m. n =15 ( sdk Δ7 , a precise excision of sdk MB05054 used as a control, and sdk MB05054 ), n =10 ( ey 3.5 -FLP, Act > CD2 > GAL4; sdk RNAi; UAS-dcr2 ), n =18 ( NP6099 > sdk RNAi; UAS-dcr2 ) and n =14 ( repo > sdk RNAi; UAS-dcr2 ). *** P
    Figure Legend Snippet: Lamina neuron placement requires sdk in photoreceptors. (A) Diagram of a coronal section through the third-instar larval brain, showing neuroblasts (nb) and lamina precursor cells (LPCs), which become postmitotic (pLPCs) behind the lamina furrow (LF; arrowhead) and differentiate into lamina neurons (LN) aligned into columns along the axons of R1-R6. Glia are shown in blue. (B-E″) Confocal images of the same view of larval brains for: (B-B″) control ( sdk MB05054 /+ ); (C-C″) sdk MB05054 ; (D-D″) sdk knockdown in the eye with ey 3.5 -FLP, Act > CD2 > GAL4 [Sdk still accumulates in LNs (asterisk)]; and (E-E″) sdk knockdown in the lamina with NP6099-GAL4 . A-E″ are labeled with anti-Sdk (B″,C″,D″,E″, green in B′,C′,D′,E′), anti-HRP to mark photoreceptor axons (red in B,B′,C,C′,D,D′,E,E′), anti-β-galactosidase (β-gal) reflecting dac-lacZ (green in B,C) or anti-Dac (green in D,E) to mark lamina neurons, anti-Repo to mark glia (blue in B,C), or anti-β-gal (blue in D,D′) or anti-GFP (blue in E,E′) to mark the domain of RNAi expression. Lamina neurons are misplaced in sdk mutants and when sdk is knocked down in the eye (empty arrowheads, C,D), but not when it is knocked down in lamina neurons or in glia with repo-GAL4 . Filled arrowheads mark the lamina furrow. (F) The number of LNs per µm in or beyond the lamina plexus (LP; arrows in A-E′) in the indicated genotypes. Data are mean±s.e.m. n =15 ( sdk Δ7 , a precise excision of sdk MB05054 used as a control, and sdk MB05054 ), n =10 ( ey 3.5 -FLP, Act > CD2 > GAL4; sdk RNAi; UAS-dcr2 ), n =18 ( NP6099 > sdk RNAi; UAS-dcr2 ) and n =14 ( repo > sdk RNAi; UAS-dcr2 ). *** P

    Techniques Used: Activated Clotting Time Assay, Labeling, Expressing

    28) Product Images from "Restrictive Expression of Acid-Sensing Ion Channel 5 (Asic5) in Unipolar Brush Cells of the Vestibulocerebellum"

    Article Title: Restrictive Expression of Acid-Sensing Ion Channel 5 (Asic5) in Unipolar Brush Cells of the Vestibulocerebellum

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0091326

    Asic5 is restrictively expressed in interneurons in the granular layer. A. Representative midsagittal section (200 µm) of the cerebellum from an Asic5 tm2a(KOMP)Wtsi mouse stained for β-galactosidase activity. B. Lobules IX and X in the (boxed) section in 3A shown at a magnified scale. C. Ten sequential sagittal sections through one half of the entire cerebellum of the reporter mouse. Sections are shown in a lateral to medial arrangement with the top left being the most lateral and the lower right (second to last) closest to the midline. Raw data are identical to that shown in 3A. Images were color and contrast manipulated to emphasize staining for β-galactosidase activity with yellow representing no staining and green representing robust staining. The final image in 3C represents a composite of the whole cerebellum collapsed into a 2-D rendering shown in the transverse plane. This rendering is a compilation of the 10 sagittal sections shown in 3B stacked lateral to medial, flipped 90° and collapsed with the left half of this figure being a mirror image of the right.
    Figure Legend Snippet: Asic5 is restrictively expressed in interneurons in the granular layer. A. Representative midsagittal section (200 µm) of the cerebellum from an Asic5 tm2a(KOMP)Wtsi mouse stained for β-galactosidase activity. B. Lobules IX and X in the (boxed) section in 3A shown at a magnified scale. C. Ten sequential sagittal sections through one half of the entire cerebellum of the reporter mouse. Sections are shown in a lateral to medial arrangement with the top left being the most lateral and the lower right (second to last) closest to the midline. Raw data are identical to that shown in 3A. Images were color and contrast manipulated to emphasize staining for β-galactosidase activity with yellow representing no staining and green representing robust staining. The final image in 3C represents a composite of the whole cerebellum collapsed into a 2-D rendering shown in the transverse plane. This rendering is a compilation of the 10 sagittal sections shown in 3B stacked lateral to medial, flipped 90° and collapsed with the left half of this figure being a mirror image of the right.

    Techniques Used: Staining, Activity Assay

    β-Gal is restrictively expressed in the vestibulocerebellum of the Asic5 tm2a(KOMP)Wtsi mouse. A. Representative images of coronal sections (200 µm) through the cerebellum and brainstem of the reporter mouse stained for β-galactosidase activity (blue). Slices are displayed in a caudal to rostral arrangement with the top being the most caudal of the three. The top, middle and bottom slices correspond with the 2 nd , 4 th and 8 th sections shown in 2B. B. Nineteen sequential coronal sections through the entire cerebellum (and part of the brainstem) of the Asic5 tm2a)KOMP)Wtsi mouse. Sections are shown in a caudal to rostral arrangement with the top left being the most caudal and the lower right (second to last) being the most rostral. Raw data are identical to that shown in 2A. Images were color and contrast manipulated to emphasize staining for β-galactosidase activity with yellow representing no staining and green representing robust staining. The final image in 2B represents a composite of the whole cerebellum (and part of the brainstem) collapsed into a 2-D rendering shown in the transverse plane. This rendering is a compilation of the 19 coronal sections shown in 2B stacked caudal to rostral, flipped 90° and collapsed.
    Figure Legend Snippet: β-Gal is restrictively expressed in the vestibulocerebellum of the Asic5 tm2a(KOMP)Wtsi mouse. A. Representative images of coronal sections (200 µm) through the cerebellum and brainstem of the reporter mouse stained for β-galactosidase activity (blue). Slices are displayed in a caudal to rostral arrangement with the top being the most caudal of the three. The top, middle and bottom slices correspond with the 2 nd , 4 th and 8 th sections shown in 2B. B. Nineteen sequential coronal sections through the entire cerebellum (and part of the brainstem) of the Asic5 tm2a)KOMP)Wtsi mouse. Sections are shown in a caudal to rostral arrangement with the top left being the most caudal and the lower right (second to last) being the most rostral. Raw data are identical to that shown in 2A. Images were color and contrast manipulated to emphasize staining for β-galactosidase activity with yellow representing no staining and green representing robust staining. The final image in 2B represents a composite of the whole cerebellum (and part of the brainstem) collapsed into a 2-D rendering shown in the transverse plane. This rendering is a compilation of the 19 coronal sections shown in 2B stacked caudal to rostral, flipped 90° and collapsed.

    Techniques Used: Staining, Activity Assay

    29) Product Images from "Changes in Sef Levels Influence Auditory Brainstem Development and Function"

    Article Title: Changes in Sef Levels Influence Auditory Brainstem Development and Function

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3477-06.2007

    Sef is expressed in cells bordering the rhombic lip. a , Dorsal (left) and transverse (right) diagrammatic views of the rhombic lip (pink) in an E12–E14 mouse embryo. The transverse view is through the plane indicated (dashed line); the position of the inner ear provides a guide for the rostrocaudal position in the hindbrain. At the level of the inner ear, the cochlear nucleus complex (purple) develops immediately adjacent to the rhombic lip. Dashed boxes outline the regions that are shown in c and d–g . 4th v, Fourth ventricle; cb, cerebellum. b , Dorsal view of an E13.5 Sef KST223 heterozygous embryo stained for β-galactosidase activity. Sef-β-geo is active in cells bordering the rim of the fourth ventricle, with intense activity (arrow) at the level of the inner ear, which has been dissected away. c , Transverse section of an E12.5 wild-type embryo hybridized with a probe to Sef . As indicated by X-gal staining, Sef is produced in a band of cells (bracket) adjacent to the rhombic lip and in a medial portion of the ventricular zone (arrow). d–f , Transverse adjacent sections through the E13.5 hindbrain hybridized with probes to Sef ( d ), Math1 ( e ), and Mafb ( f ). Sef is expressed adjacent to Math1 in the rhombic lip (arrowheads mark the lateral limit of Math1 expression) and abuts Mafb in the developing cochlear nucleus. g, h , Transverse semi-adjacent sections of an E14.5 Sef KST223 heterozygous embryo stained for β-galactosidase activity ( g ) and hybridized with a probe to Mafb ( h ). As seen at earlier stages, β-galactosidase activity is detected in the Sef -positive region adjacent to the rhombic lip (rl) (arrowhead indicates boundary). In addition, weakly stained cells are present in the developing cochlear nucleus, as determined by the expression of Mafb on a semi-adjacent section ( h ). The dashed boxes correspond to the region shown in i . i , High-power view of the region boxed in g . A stream of stained cells with elongated morphology (arrows) appears to move from the Sef -positive region into the cochlear nucleus anlage (cn). Scale bars: c–g , 50 μm; i , 10 μm.
    Figure Legend Snippet: Sef is expressed in cells bordering the rhombic lip. a , Dorsal (left) and transverse (right) diagrammatic views of the rhombic lip (pink) in an E12–E14 mouse embryo. The transverse view is through the plane indicated (dashed line); the position of the inner ear provides a guide for the rostrocaudal position in the hindbrain. At the level of the inner ear, the cochlear nucleus complex (purple) develops immediately adjacent to the rhombic lip. Dashed boxes outline the regions that are shown in c and d–g . 4th v, Fourth ventricle; cb, cerebellum. b , Dorsal view of an E13.5 Sef KST223 heterozygous embryo stained for β-galactosidase activity. Sef-β-geo is active in cells bordering the rim of the fourth ventricle, with intense activity (arrow) at the level of the inner ear, which has been dissected away. c , Transverse section of an E12.5 wild-type embryo hybridized with a probe to Sef . As indicated by X-gal staining, Sef is produced in a band of cells (bracket) adjacent to the rhombic lip and in a medial portion of the ventricular zone (arrow). d–f , Transverse adjacent sections through the E13.5 hindbrain hybridized with probes to Sef ( d ), Math1 ( e ), and Mafb ( f ). Sef is expressed adjacent to Math1 in the rhombic lip (arrowheads mark the lateral limit of Math1 expression) and abuts Mafb in the developing cochlear nucleus. g, h , Transverse semi-adjacent sections of an E14.5 Sef KST223 heterozygous embryo stained for β-galactosidase activity ( g ) and hybridized with a probe to Mafb ( h ). As seen at earlier stages, β-galactosidase activity is detected in the Sef -positive region adjacent to the rhombic lip (rl) (arrowhead indicates boundary). In addition, weakly stained cells are present in the developing cochlear nucleus, as determined by the expression of Mafb on a semi-adjacent section ( h ). The dashed boxes correspond to the region shown in i . i , High-power view of the region boxed in g . A stream of stained cells with elongated morphology (arrows) appears to move from the Sef -positive region into the cochlear nucleus anlage (cn). Scale bars: c–g , 50 μm; i , 10 μm.

    Techniques Used: Staining, Activity Assay, Produced, Expressing

    Decreased GFAP levels in cochlear nucleus astrocytes in strongly affected Sef KST223 mutants. a, a' , A coronal section through DCN from a P14 Sef KST223 heterozygote double labeled for β-galactosidase activity ( a ) and antibodies to GFAP ( a' ). Sef-β-geo and GFAP have similar patterns of distribution, with intense expression in the molecular layer (arrows). b , Double labeling of a P17 Sef KST223 heterozygote with antibodies to TuJ1 (green) and β-galactosidase (β-gal; red) shows production of Sef-β-geo in small non-neuronal cells (arrows) in the molecular layer. c, c' , Merged ( c ) and single channel ( c' ) Z series projections of two astrocytes (arrows). Sef-β-geo (red) is present in a puncate pattern within the cytoplasm of GFAP-positive astrocytes (green). β-gal, β-Galactosidase. d–g , Anti-GFAP antibody labeling of coronal sections from a wild type ( d, f ) and a Sef KST223 homozygote ( e, g ) through AVCN at the entry of the eighth nerve (8th n.). Dorsal (D) is up and to the left; lateral (L) is up and to the right. The dashed line indicates the boundary between the cochlear nucleus and the rest of the brainstem. The sections were processed and analyzed in parallel, using the same settings for image capture. High-power views of the granule cell layer are shown in f and g and correspond to the region boxed in d . In normal animals, GFAP staining is prominent in the microneuronal shell (arrows) and in the eighth nerve. Although GFAP continues to be expressed at high levels on the surface of the cochlear nucleus and in scattered cells ( g , arrows), very little expression is seen in the microneuronal shell of a Sef KST223 homozygote that had significantly elevated ABR thresholds in both ears (55 dB on the right and 80 dB on the left for wave iii). cb, Cerebellum. Scale bars: a, b, d , 40 μm; c , 5 μm; f , 10 μm.
    Figure Legend Snippet: Decreased GFAP levels in cochlear nucleus astrocytes in strongly affected Sef KST223 mutants. a, a' , A coronal section through DCN from a P14 Sef KST223 heterozygote double labeled for β-galactosidase activity ( a ) and antibodies to GFAP ( a' ). Sef-β-geo and GFAP have similar patterns of distribution, with intense expression in the molecular layer (arrows). b , Double labeling of a P17 Sef KST223 heterozygote with antibodies to TuJ1 (green) and β-galactosidase (β-gal; red) shows production of Sef-β-geo in small non-neuronal cells (arrows) in the molecular layer. c, c' , Merged ( c ) and single channel ( c' ) Z series projections of two astrocytes (arrows). Sef-β-geo (red) is present in a puncate pattern within the cytoplasm of GFAP-positive astrocytes (green). β-gal, β-Galactosidase. d–g , Anti-GFAP antibody labeling of coronal sections from a wild type ( d, f ) and a Sef KST223 homozygote ( e, g ) through AVCN at the entry of the eighth nerve (8th n.). Dorsal (D) is up and to the left; lateral (L) is up and to the right. The dashed line indicates the boundary between the cochlear nucleus and the rest of the brainstem. The sections were processed and analyzed in parallel, using the same settings for image capture. High-power views of the granule cell layer are shown in f and g and correspond to the region boxed in d . In normal animals, GFAP staining is prominent in the microneuronal shell (arrows) and in the eighth nerve. Although GFAP continues to be expressed at high levels on the surface of the cochlear nucleus and in scattered cells ( g , arrows), very little expression is seen in the microneuronal shell of a Sef KST223 homozygote that had significantly elevated ABR thresholds in both ears (55 dB on the right and 80 dB on the left for wave iii). cb, Cerebellum. Scale bars: a, b, d , 40 μm; c , 5 μm; f , 10 μm.

    Techniques Used: Labeling, Activity Assay, Expressing, Antibody Labeling, Staining

    30) Product Images from "Loss of a proteostatic checkpoint in intestinal stem cells contributes to age-related epithelial dysfunction"

    Article Title: Loss of a proteostatic checkpoint in intestinal stem cells contributes to age-related epithelial dysfunction

    Journal: Nature Communications

    doi: 10.1038/s41467-019-08982-9

    Protein aggregates induce a transient cell cycle arrest in ISCs. a , b GFP-marked lineages (green) from Rpn3 RNAi expressing ISCs in 8-day-old flies. Immunohistochemistry: a anti-poly-ubiquitin (white, left; black, right), b anti-Delta (white). b 1-way ANOVA / Dunnett’s ( n = 41, 73, 66, and 25 clones from 3, 4, 4, 3 independent animals respectively). c mRFP-Htt Q138 expressed for 24 h in ISCs of 11-day-old flies. Delta (Dl; white). Graph: RFP signal normalized to background. Two-tailed unpaired t-test: ( n = 75 and 57 Dl+ cells from five and three animals respectively, from two independent experiments). d ISC lineages from 4 days old flies labelled with β-galactosidase (βgal, green) using Flp-mediated somatic recombination of a split lacZ gene (X-15-33/x-15-29 47 ), carrying or not mRFP-Htt Q138 (RU486-inducible ISC/EB driver 5961::GS). Beta-galactosidase (green), RFP-labeled aggregates red. Yellow arrowheads point out βgal-labeled ISCs. 1-way ANOVA/Sidak’s ( n = 76, 154, 72, and 97 clones from 10, 13, 9, and 9 animals respectively, from two independent experiments). a – d , Representative areas of posterior midgut. DNA (Hoechst, blue); scale bars 20 µm and 10 µm ( c , inset). e Quantification of phospho-Histone H3 (pH3)+ cells after 24 h expression of mRFP-Htt Q138 in ISCs of 6 to 8-day-old flies during infection with Ecc15 (or Mock, M; first graph), or followed by 24 h Ecc15 immediately (second graph), or by 8 h Ecc15 one week (third graph), or two weeks (fourth graph) later. Two-way ANOVA with Tukey’s ( n = 12, 15, 20, 19 guts (first); 11, 16, 26, 24 guts (second); 12, 14, 19, 20 guts (third); 5, 6, 13, 13 guts (fourth graph)). f pH3+ cell quantification in 12-day-old flies after 5 mM Paraquat (PQ) or Mock (M) treatment at 25 °C, followed by regular food at 29 °C for 12 h. Two-way ANOVA with Sidak’s ( n = 32, 32, 40, 15 guts, from two independent experiments). g Survival of 12-day-old flies fed 5 mM PQ or Mock at 25 °C for 24 h, followed by regular food at 29 °C for 24 h ( n = 3 biological replicates). Two-way ANOVA with Tukey’s. Means and s.e.m. shown in all graphs; ns, not significant, **** P
    Figure Legend Snippet: Protein aggregates induce a transient cell cycle arrest in ISCs. a , b GFP-marked lineages (green) from Rpn3 RNAi expressing ISCs in 8-day-old flies. Immunohistochemistry: a anti-poly-ubiquitin (white, left; black, right), b anti-Delta (white). b 1-way ANOVA / Dunnett’s ( n = 41, 73, 66, and 25 clones from 3, 4, 4, 3 independent animals respectively). c mRFP-Htt Q138 expressed for 24 h in ISCs of 11-day-old flies. Delta (Dl; white). Graph: RFP signal normalized to background. Two-tailed unpaired t-test: ( n = 75 and 57 Dl+ cells from five and three animals respectively, from two independent experiments). d ISC lineages from 4 days old flies labelled with β-galactosidase (βgal, green) using Flp-mediated somatic recombination of a split lacZ gene (X-15-33/x-15-29 47 ), carrying or not mRFP-Htt Q138 (RU486-inducible ISC/EB driver 5961::GS). Beta-galactosidase (green), RFP-labeled aggregates red. Yellow arrowheads point out βgal-labeled ISCs. 1-way ANOVA/Sidak’s ( n = 76, 154, 72, and 97 clones from 10, 13, 9, and 9 animals respectively, from two independent experiments). a – d , Representative areas of posterior midgut. DNA (Hoechst, blue); scale bars 20 µm and 10 µm ( c , inset). e Quantification of phospho-Histone H3 (pH3)+ cells after 24 h expression of mRFP-Htt Q138 in ISCs of 6 to 8-day-old flies during infection with Ecc15 (or Mock, M; first graph), or followed by 24 h Ecc15 immediately (second graph), or by 8 h Ecc15 one week (third graph), or two weeks (fourth graph) later. Two-way ANOVA with Tukey’s ( n = 12, 15, 20, 19 guts (first); 11, 16, 26, 24 guts (second); 12, 14, 19, 20 guts (third); 5, 6, 13, 13 guts (fourth graph)). f pH3+ cell quantification in 12-day-old flies after 5 mM Paraquat (PQ) or Mock (M) treatment at 25 °C, followed by regular food at 29 °C for 12 h. Two-way ANOVA with Sidak’s ( n = 32, 32, 40, 15 guts, from two independent experiments). g Survival of 12-day-old flies fed 5 mM PQ or Mock at 25 °C for 24 h, followed by regular food at 29 °C for 24 h ( n = 3 biological replicates). Two-way ANOVA with Tukey’s. Means and s.e.m. shown in all graphs; ns, not significant, **** P

    Techniques Used: Expressing, Immunohistochemistry, Clone Assay, Two Tailed Test, Labeling, Infection

    31) Product Images from "Biodistribution and retargeting of FX-binding ablated adenovirus serotype 5 vectors"

    Article Title: Biodistribution and retargeting of FX-binding ablated adenovirus serotype 5 vectors

    Journal: Blood

    doi: 10.1182/blood-2009-12-260026

    Transduction profiles of Ad5, Ad5-HVR7(Ad26), and Ad5-HVR5*7*E451Q in control (CL−) or macrophage-depleted (CL+) mice . β-galactosidase expression was quantified by ELISA and normalized to total protein content. Animals were killed 48 hours
    Figure Legend Snippet: Transduction profiles of Ad5, Ad5-HVR7(Ad26), and Ad5-HVR5*7*E451Q in control (CL−) or macrophage-depleted (CL+) mice . β-galactosidase expression was quantified by ELISA and normalized to total protein content. Animals were killed 48 hours

    Techniques Used: Transduction, Mouse Assay, Expressing, Enzyme-linked Immunosorbent Assay

    Liver transduction of hexon modified Ad5 vectors using 1 × 10 11 vp per mouse . β-galactosidase transgene expression of Ad5 and hexon modified adenoviral vectors after high-dose administration of Ad to macrophage-depleted mice. Animals were
    Figure Legend Snippet: Liver transduction of hexon modified Ad5 vectors using 1 × 10 11 vp per mouse . β-galactosidase transgene expression of Ad5 and hexon modified adenoviral vectors after high-dose administration of Ad to macrophage-depleted mice. Animals were

    Techniques Used: Transduction, Modification, Expressing, Mouse Assay

    32) Product Images from "Pre-Existing Mature Oligodendrocytes Do Not Contribute to Remyelination following Toxin-Induced Spinal Cord Demyelination"

    Article Title: Pre-Existing Mature Oligodendrocytes Do Not Contribute to Remyelination following Toxin-Induced Spinal Cord Demyelination

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.11.005

    A: Five days postlesion (dpl) in Opalin-iCreER T2 : Tau-mGfp-LacZ mouse: Ki-67–Green fluorescent protein (Gfp)-positive oligodendrocyte (OL) from the presumed lesion edge surrounded by Ki-67 + cells. B: Number of membranous Gfp + cells in contralateral
    Figure Legend Snippet: A: Five days postlesion (dpl) in Opalin-iCreER T2 : Tau-mGfp-LacZ mouse: Ki-67–Green fluorescent protein (Gfp)-positive oligodendrocyte (OL) from the presumed lesion edge surrounded by Ki-67 + cells. B: Number of membranous Gfp + cells in contralateral

    Techniques Used:

    33) Product Images from "Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes"

    Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

    Journal: PLoS Biology

    doi: 10.1371/journal.pbio.1000283

    GRASP-1 is required for the maintenance of dendritic spines. (A) Representative high magnification images of dendrites of hippocampal neurons co-transfected at DIV13 for 4 d with β-galactosidase (to mark the dendrites), and either pSuper, pSuper-GRASP-1-shRNA#2, GRASP-1-shRNA#2 and GFP-GRASP-1*, Rab4S22N or Rab11S25N, and labeled with anti-β-galactosidase. (B) Quantification of number of protrusions per 10 µm dendrites in hippocampal neurons transfected as indicated in (A). (C) Percentage of spines of hippocampal neurons transfected as indicated in (A). (D) Neurons expressing GFP (to mark the dendrite), and either pSuper or pSuper-GRASP-1-shRNA#2 were stimulated with glycine (200 mM, 3 min), and then imaged for > 30 min after glycine stimulation. Arrows indicated spine formation. Closed and open arrowheads represent spine growth and stable protrusions, respectively. (E) Quantification of protrusion formation (top) and spine growth (bottom) following glycine stimulation. N, number of dendritic protrusions per 10 µm at the indicated time; N 0 , average number of dendritic protrusions per 10 µm before application of glycine. Spine growth was probed as the change in sum of spine widths per 10 µm and comprises both addition of new spines and growth of pre-existing spines. Glycine-stimulated spine growth is blocked by GRASP-1-shRNA#2 (bottom). (F) High magnification images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-GRASP-1 (red) and GFP-TfR. (G,H) Percentage of spines containing TfR-GFP positive endosomes at the indicated locations. Hippocampal neurons were co-transfected at DIV13 for 4 d with β-galactosidase (to mark dendrites) and GFP-TfR (to mark endosomes) and pSuper control vector or pSuper-GRASP-1-shRNA#2 as shown in (H). Closed and open arrowheads denote protrusions with and without GFP-TfR marked endosomes in the spine head, respectively. Error bars indicate S.E.M. ** p
    Figure Legend Snippet: GRASP-1 is required for the maintenance of dendritic spines. (A) Representative high magnification images of dendrites of hippocampal neurons co-transfected at DIV13 for 4 d with β-galactosidase (to mark the dendrites), and either pSuper, pSuper-GRASP-1-shRNA#2, GRASP-1-shRNA#2 and GFP-GRASP-1*, Rab4S22N or Rab11S25N, and labeled with anti-β-galactosidase. (B) Quantification of number of protrusions per 10 µm dendrites in hippocampal neurons transfected as indicated in (A). (C) Percentage of spines of hippocampal neurons transfected as indicated in (A). (D) Neurons expressing GFP (to mark the dendrite), and either pSuper or pSuper-GRASP-1-shRNA#2 were stimulated with glycine (200 mM, 3 min), and then imaged for > 30 min after glycine stimulation. Arrows indicated spine formation. Closed and open arrowheads represent spine growth and stable protrusions, respectively. (E) Quantification of protrusion formation (top) and spine growth (bottom) following glycine stimulation. N, number of dendritic protrusions per 10 µm at the indicated time; N 0 , average number of dendritic protrusions per 10 µm before application of glycine. Spine growth was probed as the change in sum of spine widths per 10 µm and comprises both addition of new spines and growth of pre-existing spines. Glycine-stimulated spine growth is blocked by GRASP-1-shRNA#2 (bottom). (F) High magnification images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-GRASP-1 (red) and GFP-TfR. (G,H) Percentage of spines containing TfR-GFP positive endosomes at the indicated locations. Hippocampal neurons were co-transfected at DIV13 for 4 d with β-galactosidase (to mark dendrites) and GFP-TfR (to mark endosomes) and pSuper control vector or pSuper-GRASP-1-shRNA#2 as shown in (H). Closed and open arrowheads denote protrusions with and without GFP-TfR marked endosomes in the spine head, respectively. Error bars indicate S.E.M. ** p

    Techniques Used: Transfection, shRNA, Labeling, Expressing, Plasmid Preparation

    34) Product Images from "Endothelial Wnt/?-catenin signaling inhibits glioma angiogenesis and normalizes tumor blood vessels by inducing PDGF-B expression"

    Article Title: Endothelial Wnt/?-catenin signaling inhibits glioma angiogenesis and normalizes tumor blood vessels by inducing PDGF-B expression

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20111580

    Vascular effects observed in Wnt1-expressing gliomas are EC autonomous. (A, left) IF staining for β-galactosidase (β-gal), Podxl, and TOPRO-3 on GL261 glioma sections from Pdgfb-iCreERT2 × ROSA26 STOPfloxLacZ mice treated with tamoxifen. (right) Quantification of β-gal + vessels. (B, left) IF staining for Podxl and TOPRO-3 on GL261 glioma sections from Pdgfb-iCreERT2 (control) and Pdgfb-iCreERT2 × βCat Ex3flox (GOF) mice. (right) Quantification of Podxl + areas within tumors ( n = 4 tumors/group, 10 pictures/tumor; *, P
    Figure Legend Snippet: Vascular effects observed in Wnt1-expressing gliomas are EC autonomous. (A, left) IF staining for β-galactosidase (β-gal), Podxl, and TOPRO-3 on GL261 glioma sections from Pdgfb-iCreERT2 × ROSA26 STOPfloxLacZ mice treated with tamoxifen. (right) Quantification of β-gal + vessels. (B, left) IF staining for Podxl and TOPRO-3 on GL261 glioma sections from Pdgfb-iCreERT2 (control) and Pdgfb-iCreERT2 × βCat Ex3flox (GOF) mice. (right) Quantification of Podxl + areas within tumors ( n = 4 tumors/group, 10 pictures/tumor; *, P

    Techniques Used: Expressing, Staining, Mouse Assay

    35) Product Images from "Identification of Epithelial to Mesenchymal Transition as a Novel Source of Fibroblasts in Intestinal Fibrosis *"

    Article Title: Identification of Epithelial to Mesenchymal Transition as a Novel Source of Fibroblasts in Intestinal Fibrosis *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.102012

    Evidence for EMT associated with intestinal fibrosis in Villin-LacZ mice. Frozen colonic tissue sections from ethanol control and TNBS-treated Villin-LacZ mice were either labeled with antibodies to β-gal ( red ) and FSP1 ( green ) or with antibodies
    Figure Legend Snippet: Evidence for EMT associated with intestinal fibrosis in Villin-LacZ mice. Frozen colonic tissue sections from ethanol control and TNBS-treated Villin-LacZ mice were either labeled with antibodies to β-gal ( red ) and FSP1 ( green ) or with antibodies

    Techniques Used: Mouse Assay, Labeling

    36) Product Images from "Drosophila ATP6AP2/VhaPRR functions both as a novel planar cell polarity core protein and a regulator of endosomal trafficking"

    Article Title: Drosophila ATP6AP2/VhaPRR functions both as a novel planar cell polarity core protein and a regulator of endosomal trafficking

    Journal: The EMBO Journal

    doi: 10.1038/emboj.2012.323

    Loss of VhaPRR causes PCP defects and PCP core protein mislocalization. All clones are marked by loss of β-galactosidase (β-gal; only shown in ( F′ , G′ ) in red). Clone outlines are marked with a dotted white line. In all
    Figure Legend Snippet: Loss of VhaPRR causes PCP defects and PCP core protein mislocalization. All clones are marked by loss of β-galactosidase (β-gal; only shown in ( F′ , G′ ) in red). Clone outlines are marked with a dotted white line. In all

    Techniques Used: Clone Assay

    Lack of VhaPRR causes cell packing defects and increases junctional E-Cadherin. ( A , A′ ) E-Cadherin (red) levels are elevated in VhaPRR mutant clones at 32 h APF. Lack of β-gal (blue in A′ ) marks the clone region. Hexagonal packing
    Figure Legend Snippet: Lack of VhaPRR causes cell packing defects and increases junctional E-Cadherin. ( A , A′ ) E-Cadherin (red) levels are elevated in VhaPRR mutant clones at 32 h APF. Lack of β-gal (blue in A′ ) marks the clone region. Hexagonal packing

    Techniques Used: Mutagenesis, Clone Assay

    37) Product Images from "A Differentially Autoregulated Pet-1 Enhancer Region Is a Critical Target of the Transcriptional Cascade That Governs Serotonin Neuron Development"

    Article Title: A Differentially Autoregulated Pet-1 Enhancer Region Is a Critical Target of the Transcriptional Cascade That Governs Serotonin Neuron Development

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4979-04.2005

    Pet-1 enhancer activity is autoregulated in a subset of 5-HT neurons. A - F , Coimmunostaining ( C , F ) of 5-HT ( A , D ) with β-galactosidase ( B , E ) in 20 μm sagittal sections of Pet-1 +/- ( A-C ) and Pet-1 -/- ( D-F ) E18 rostral hindbrain. 5-HT/β-gal coimmunostaining of Pet-1 +/- ( G ) and Pet-1 -/- ( H ) postnatal day 3 midbrain. F , H , Asterisks indicate representative lacZ - /5-HT + and lacZ + /5-HT - cells in the Pet-1 -/- embryo ( F ) and neonate ( H ). Arrowheads indicate cells coexpressing 540Z and 5-HT in the Pet-1 -/- embryo ( F ) and neonate ( H ). Scale bars: A - F , 100 μm; G , H , 50 μm.
    Figure Legend Snippet: Pet-1 enhancer activity is autoregulated in a subset of 5-HT neurons. A - F , Coimmunostaining ( C , F ) of 5-HT ( A , D ) with β-galactosidase ( B , E ) in 20 μm sagittal sections of Pet-1 +/- ( A-C ) and Pet-1 -/- ( D-F ) E18 rostral hindbrain. 5-HT/β-gal coimmunostaining of Pet-1 +/- ( G ) and Pet-1 -/- ( H ) postnatal day 3 midbrain. F , H , Asterisks indicate representative lacZ - /5-HT + and lacZ + /5-HT - cells in the Pet-1 -/- embryo ( F ) and neonate ( H ). Arrowheads indicate cells coexpressing 540Z and 5-HT in the Pet-1 -/- embryo ( F ) and neonate ( H ). Scale bars: A - F , 100 μm; G , H , 50 μm.

    Techniques Used: Positron Emission Tomography, Activity Assay

    540Z expression recapitulates the temporal expression of the endogenous Pet-1 gene. Embryonic sagittal sections (10 μm) were used in all of the panels. A-K , Colocalization ( C , F-J ) of 5-HT ( B , E ) with β-galactosidase ( A , D ) at embryonic ages E11.5 ( A-C ), E12 ( D-F ), E13.5 ( G , H , K ), and E16 ( I , J ). K , β-galactosidase expression in whole-embryo view. R, Rostral domain; C, caudal domain; A, anterior; V, ventral; P, posterior; D, dorsal. Scale bars: A-F , I , J , 100 μm; G , H , 50 μm; K , 250 μm. Arrows ( D , F ) and asterisks ( G , H ) identify lacZ + /5-HT - neurons. Orientation of the section as identified for C applies to all of the panels.
    Figure Legend Snippet: 540Z expression recapitulates the temporal expression of the endogenous Pet-1 gene. Embryonic sagittal sections (10 μm) were used in all of the panels. A-K , Colocalization ( C , F-J ) of 5-HT ( B , E ) with β-galactosidase ( A , D ) at embryonic ages E11.5 ( A-C ), E12 ( D-F ), E13.5 ( G , H , K ), and E16 ( I , J ). K , β-galactosidase expression in whole-embryo view. R, Rostral domain; C, caudal domain; A, anterior; V, ventral; P, posterior; D, dorsal. Scale bars: A-F , I , J , 100 μm; G , H , 50 μm; K , 250 μm. Arrows ( D , F ) and asterisks ( G , H ) identify lacZ + /5-HT - neurons. Orientation of the section as identified for C applies to all of the panels.

    Techniques Used: Expressing, Positron Emission Tomography

    38) Product Images from "Insights into Hox Protein Function from a Large Scale Combinatorial Analysis of Protein Domains"

    Article Title: Insights into Hox Protein Function from a Large Scale Combinatorial Analysis of Protein Domains

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1002302

    Mutually suppressive interaction of protein domains. A. Thoracic restricted expression of Dll (white arrows) followed by Dll enhancer driven β-gal (green) results from repression by AbdA (red) in the abdomen (upper panel). Ubiquitous AbdA expression driven by arm-Gal4 represses Dll thoracic expression (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Right panels are magnification of boxed thoracic areas. B. Increased thoracic Antp expression (green, white arrows) results from AbdA (red) repression in the abdomen (upper panels). Ubiquitous AbdA expression driven by arm-Gal4 represses Antp expression in the thorax (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Right panels are magnification of boxed thoracic areas. C. Abdominal segments are characterized by refringent denticles organized in a trapezoidal shape in segments A2 but not A1, while T2/T3 thoracic segments harbors thinner denticles (left panel). Upon AbdA thoracic expression driven by arm-Gal4 , the first abdominal segment A1 and thoracic segments acquire abdominal features, including abdominal type of denticles, trapezoidal organization of denticles and suppression of a T1 specific feature (white arrow), the “beard” (middle panel). Full or intermediate transformations were observed for AbdA variants (see Text S1 for quantifying criteria). The effect of the AbdA HX/TD variants is illustrated (right panel). Weak A1 (wA1) stands for a transformation of thoracic denticles toward abdominal type of denticles, with an organization typical of A1, but with only a partial suppression of the beard in T1 (arrow). D. Snapshots from movies illustrating locomotion in wild type larvae (left panels), or in larvae expressing ubiquitously AbdA (middle panels) or AbdA HX/UA variant (right panels) driven by the arm-Gal4 driver. White boxed areas show the progression of a peristaltic waves in the abdomen. The red boxed area shows an ectopic peristaltic wave in the thorax following ectopic AbdA expression in the thorax. Graphs in A–D (% of remaining activities compared to the wild type AbdA protein (WT) following domain mutations) using the boxplot representation summarize quantitative analyses (see Text S1 and Figure S7 ( Dll ), S8 (Antp), and S9 (A2 epidermal morphology) for full illustration, and Figure S10 for data on larval locomotion experiments. A graded color-coded bar above the graphs illustrates the level of protein activity, ranging from light green (full activity) to black (no activity).
    Figure Legend Snippet: Mutually suppressive interaction of protein domains. A. Thoracic restricted expression of Dll (white arrows) followed by Dll enhancer driven β-gal (green) results from repression by AbdA (red) in the abdomen (upper panel). Ubiquitous AbdA expression driven by arm-Gal4 represses Dll thoracic expression (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Right panels are magnification of boxed thoracic areas. B. Increased thoracic Antp expression (green, white arrows) results from AbdA (red) repression in the abdomen (upper panels). Ubiquitous AbdA expression driven by arm-Gal4 represses Antp expression in the thorax (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Right panels are magnification of boxed thoracic areas. C. Abdominal segments are characterized by refringent denticles organized in a trapezoidal shape in segments A2 but not A1, while T2/T3 thoracic segments harbors thinner denticles (left panel). Upon AbdA thoracic expression driven by arm-Gal4 , the first abdominal segment A1 and thoracic segments acquire abdominal features, including abdominal type of denticles, trapezoidal organization of denticles and suppression of a T1 specific feature (white arrow), the “beard” (middle panel). Full or intermediate transformations were observed for AbdA variants (see Text S1 for quantifying criteria). The effect of the AbdA HX/TD variants is illustrated (right panel). Weak A1 (wA1) stands for a transformation of thoracic denticles toward abdominal type of denticles, with an organization typical of A1, but with only a partial suppression of the beard in T1 (arrow). D. Snapshots from movies illustrating locomotion in wild type larvae (left panels), or in larvae expressing ubiquitously AbdA (middle panels) or AbdA HX/UA variant (right panels) driven by the arm-Gal4 driver. White boxed areas show the progression of a peristaltic waves in the abdomen. The red boxed area shows an ectopic peristaltic wave in the thorax following ectopic AbdA expression in the thorax. Graphs in A–D (% of remaining activities compared to the wild type AbdA protein (WT) following domain mutations) using the boxplot representation summarize quantitative analyses (see Text S1 and Figure S7 ( Dll ), S8 (Antp), and S9 (A2 epidermal morphology) for full illustration, and Figure S10 for data on larval locomotion experiments. A graded color-coded bar above the graphs illustrates the level of protein activity, ranging from light green (full activity) to black (no activity).

    Techniques Used: Expressing, Transformation Assay, Variant Assay, Activity Assay

    Additive contribution of protein domains. Oenocytes ventrally located in the abdomen are visualized by β-gal (green) driven by the seven-up ( svp ) promoter (white arrows, upper panel). Expression of AbdA (red) in the thorax through the arm-Gal4 driver induces ectopic oenocytes in the thorax (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Boxed areas highlight thoracic segments. Right panels are magnifications of the boxed areas. Graphs (% of remaining activities compared to the wild type AbdA protein (WT) following domain mutations) using the boxplot representation on the right summarize quantitative analyses (see Text S1 and Figure S4 for full illustration). A graded color-coded bar above the graphs illustrates the level of protein activity, ranging from light green (full activity) to black (no activity).
    Figure Legend Snippet: Additive contribution of protein domains. Oenocytes ventrally located in the abdomen are visualized by β-gal (green) driven by the seven-up ( svp ) promoter (white arrows, upper panel). Expression of AbdA (red) in the thorax through the arm-Gal4 driver induces ectopic oenocytes in the thorax (red arrows, middle panels). The effect of the AbdA HX/UA variants is illustrated (lower panels). Boxed areas highlight thoracic segments. Right panels are magnifications of the boxed areas. Graphs (% of remaining activities compared to the wild type AbdA protein (WT) following domain mutations) using the boxplot representation on the right summarize quantitative analyses (see Text S1 and Figure S4 for full illustration). A graded color-coded bar above the graphs illustrates the level of protein activity, ranging from light green (full activity) to black (no activity).

    Techniques Used: Expressing, Activity Assay

    39) Product Images from "A late requirement for Wnt and FGF signaling during activin-induced formation of foregut endoderm from mouse embryonic stem cells"

    Article Title: A late requirement for Wnt and FGF signaling during activin-induced formation of foregut endoderm from mouse embryonic stem cells

    Journal: Developmental biology

    doi: 10.1016/j.ydbio.2009.03.026

    ES cell-derived endoderm acquire foregut cell fates ( A–J’ and P–Z’ ) OS25 (Sox2 β geo /+ ) or ( K–O ) Pdx1 LacZ /+ cells were cultured for 5 days in 100 ng/ml activin and subsequently treated with 5 ng/ml Wnt3a, 10 ng/ml FGF4 and/or 0,1 µM RA for 3 days as indicated. The co-expression of Foxa2 and Sox2 (foregut), Pdx1 (midgut) or Cdx2 (hindgut) was analysed by immunofluorescence. The expression of Sox2 ( A–E ) and Pdx1 ( K–O ) was confirmed by analysing β-galactosidase activity using X-gal staining.
    Figure Legend Snippet: ES cell-derived endoderm acquire foregut cell fates ( A–J’ and P–Z’ ) OS25 (Sox2 β geo /+ ) or ( K–O ) Pdx1 LacZ /+ cells were cultured for 5 days in 100 ng/ml activin and subsequently treated with 5 ng/ml Wnt3a, 10 ng/ml FGF4 and/or 0,1 µM RA for 3 days as indicated. The co-expression of Foxa2 and Sox2 (foregut), Pdx1 (midgut) or Cdx2 (hindgut) was analysed by immunofluorescence. The expression of Sox2 ( A–E ) and Pdx1 ( K–O ) was confirmed by analysing β-galactosidase activity using X-gal staining.

    Techniques Used: Derivative Assay, Cell Culture, Expressing, Immunofluorescence, Activity Assay, Staining

    BMP4 but not Wnt3a inhibits the expression of Foxa2 and E-cadherin, and promotes expression of Flk1 in the presence of activin The expression of Foxa2, E-cad (Cdh1), and Flk1 (β-gal) was analysed by immunofluorescence in Flk1 LacZ /+ ES cells cultured for 5 days in media containing 0, 3 or 100 ng/ml activin, 100 ng/ml Wnt3a, 10 ng/ml BMP4, 100 ng/ml activin+100 ng/ml Wnt3a, or 100 ng/ml activin+10 ng/ml BMP4.
    Figure Legend Snippet: BMP4 but not Wnt3a inhibits the expression of Foxa2 and E-cadherin, and promotes expression of Flk1 in the presence of activin The expression of Foxa2, E-cad (Cdh1), and Flk1 (β-gal) was analysed by immunofluorescence in Flk1 LacZ /+ ES cells cultured for 5 days in media containing 0, 3 or 100 ng/ml activin, 100 ng/ml Wnt3a, 10 ng/ml BMP4, 100 ng/ml activin+100 ng/ml Wnt3a, or 100 ng/ml activin+10 ng/ml BMP4.

    Techniques Used: Expressing, Immunofluorescence, Cell Culture

    40) Product Images from "dMyc is required in retinal progenitors to prevent JNK-mediated retinal glial activation"

    Article Title: dMyc is required in retinal progenitors to prevent JNK-mediated retinal glial activation

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1006647

    dMyc knockdown in the eye imaginal disc induces glia overmigration. (A) Schematic of the L2 (top left) and L3 (bottom) eye imaginal disc with color-coded expression domains of the Gal4 drivers used in this work. Red: repo -Gal4; Blue: ey -Gal4; Pink: dppblk -Gal4; Yellow: hth -Gal4. A dashed line represents the Morphogenetic Furrow (MF). (B–C) Atonal expression assessed by the reporter ato- GFP in control (B) and ey > dMyc RNAi (C). (D–E) Transverse view of the eye imaginal disc showing glia nuclei (red) and photoreceptor axons (grey) in Control (D) and ey > dMyc RNAi (E). An arrowhead indicates MF. (F–G) Photoreceptor cells stained with Elav (neuronal marker) in control (F) and ey > dMyc RNAi (G). (H–I) Wrapping glial cells are labelled with β-galactosidase to detect sprouty -LacZ (Spy-Z) (green) in control (H) and ey > dMyc RNAi (I). Cyan dashed line represents the glia overmigration position. (J–O) Glial cell membranes were detected with repo LexA-LexAopCD2-GFP (green) in control (J–L) and ey > dMyc RNAi (M–O). J’, J”, M’ and M” are magnifications of the white inset shown in panel J and M respectively. K and N correspond to transversal section of the optic stalk where wrapped axons are visible. Arrows point towards region of wrapping glia. L and O are magnifications of the dashed inset shown in panel J and M, respectively, showing glia morphology at the edge of migration. (P–Q) Early L3 eye imaginal disc of control (P) and ey > dMyc RNAi (Q). Glial cells migrate before the onset of differentiation (shown by Hrp staining) in ey > dMyc RNAi. (R) Pi3K92E knockdown in the eye disc reduces tissue growth but does not affect glia overmigration. (S) hth > dMyc RNAi eye discs showing glia overmigration. (T) repo > dcr-2 > dMyc RNAi eye discs have reduced numbers of glial cells. Glial cells stained with Repo are shown in red; Hrp or Elav are used to label photoreceptors in grey, and DAPI stains DNA in blue. A dashed line represents the MF. Scale bars correspond to 10 μm.
    Figure Legend Snippet: dMyc knockdown in the eye imaginal disc induces glia overmigration. (A) Schematic of the L2 (top left) and L3 (bottom) eye imaginal disc with color-coded expression domains of the Gal4 drivers used in this work. Red: repo -Gal4; Blue: ey -Gal4; Pink: dppblk -Gal4; Yellow: hth -Gal4. A dashed line represents the Morphogenetic Furrow (MF). (B–C) Atonal expression assessed by the reporter ato- GFP in control (B) and ey > dMyc RNAi (C). (D–E) Transverse view of the eye imaginal disc showing glia nuclei (red) and photoreceptor axons (grey) in Control (D) and ey > dMyc RNAi (E). An arrowhead indicates MF. (F–G) Photoreceptor cells stained with Elav (neuronal marker) in control (F) and ey > dMyc RNAi (G). (H–I) Wrapping glial cells are labelled with β-galactosidase to detect sprouty -LacZ (Spy-Z) (green) in control (H) and ey > dMyc RNAi (I). Cyan dashed line represents the glia overmigration position. (J–O) Glial cell membranes were detected with repo LexA-LexAopCD2-GFP (green) in control (J–L) and ey > dMyc RNAi (M–O). J’, J”, M’ and M” are magnifications of the white inset shown in panel J and M respectively. K and N correspond to transversal section of the optic stalk where wrapped axons are visible. Arrows point towards region of wrapping glia. L and O are magnifications of the dashed inset shown in panel J and M, respectively, showing glia morphology at the edge of migration. (P–Q) Early L3 eye imaginal disc of control (P) and ey > dMyc RNAi (Q). Glial cells migrate before the onset of differentiation (shown by Hrp staining) in ey > dMyc RNAi. (R) Pi3K92E knockdown in the eye disc reduces tissue growth but does not affect glia overmigration. (S) hth > dMyc RNAi eye discs showing glia overmigration. (T) repo > dcr-2 > dMyc RNAi eye discs have reduced numbers of glial cells. Glial cells stained with Repo are shown in red; Hrp or Elav are used to label photoreceptors in grey, and DAPI stains DNA in blue. A dashed line represents the MF. Scale bars correspond to 10 μm.

    Techniques Used: Expressing, Staining, Marker, Migration

    dMyc is required to prevent ectopic JNK pathway activation. (A–B)– puc E69 expression (β-galactosidase reporter for puc ; green) in control (A) and ey > dMyc RNAi (B). (C–D)–TRE-GFP expression (green) in control (C) and ey > dMyc RNAi (D). (E–H)–pJNK expression (green) in control (E and F) and ey > dMyc RNAi (G and H). A”,B”, C”,D”, F, F’, H and H’ show transversal views from the eye disc. Arrows point towards eye disc areas with high JNK pathway activation and asterisks represent JNK activation in glia. Glial cells stained with repo are shown in red, Hrp shows the photoreceptors in grey and DAPI stains DNA in blue. A yellow dashed line or arrowhead represents the MF. Scale bars correspond to 10 μm.
    Figure Legend Snippet: dMyc is required to prevent ectopic JNK pathway activation. (A–B)– puc E69 expression (β-galactosidase reporter for puc ; green) in control (A) and ey > dMyc RNAi (B). (C–D)–TRE-GFP expression (green) in control (C) and ey > dMyc RNAi (D). (E–H)–pJNK expression (green) in control (E and F) and ey > dMyc RNAi (G and H). A”,B”, C”,D”, F, F’, H and H’ show transversal views from the eye disc. Arrows point towards eye disc areas with high JNK pathway activation and asterisks represent JNK activation in glia. Glial cells stained with repo are shown in red, Hrp shows the photoreceptors in grey and DAPI stains DNA in blue. A yellow dashed line or arrowhead represents the MF. Scale bars correspond to 10 μm.

    Techniques Used: Activation Assay, Expressing, Staining

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    Article Snippet: .. Sections at 12 µm were generated from fixed brain tissue and immunohistochemistry was performed as described in Zarbalis et al. [ ] using the following antibodies: mouse anti-Col1a1 1:100 (Sigma); rabbit anti-Col1 1:100 (Abcam); rabbit anti-PDGFrβ 1:200 (Cell Signaling Technology); rat anti-PDGFrβ 1:200 (Novus Biologicals); rat anti-PDGFrα 1:200 (BD Bioscience); mouse anti-CoupTF2 1:300 (R & D); rabbit anti-Raldh1 1:200 (Abcam); rabbit anti-Raldh2 1:400 (Sigma-Aldrich); mouse anti-CRABP1 1:100 (Abcam); rabbit anti-CRABP2 1:100 (Proteintech); mouse CD68 1:500 (Dako); rabbit anti-β-galactosidase 1:500 (MP Biomed); chicken anti-β-galactosidase 1:500 (Abcam); mouse anti-NeuN 1:500 (Millipore); rabbit anti-glial fibrillary acidic protein (GFAP) 1:500 (Sigma-Aldrich). .. Following incubation with primary antibodies, sections were incubated with appropriate Alexafluor-conjugated secondary antibodies (Invitrogen) at 1:500, Alexafluor 633-conjugated isolectin-B4 (Invitrogen) at 1:500 to label blood vessels and microglia [ ], and DAPI (4′,6-diamidino-2-phenylindole) to label nuclei (Invitrogen).

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    Generated:

    Article Title: Col1a1+ perivascular cells in the brain are a source of retinoic acid following stroke
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    Western Blot:

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