Journal: Developmental biology

Article Title: Oxygen regulates vascular endothelial growth factor-mediated vasculogenesis and tubulogenesis.

doi: 10.1006/dbio.1997.8513

Figure Lengend Snippet: FIG. 8. (A) 3% O2-treated explant showing immunoreactive Flt-1 localized to endothelial cells and tubular epithelial cells (original magnification, 150). (B) 3% O2-treated explant showing immunoreactive Flk-1 localized to endothelial cells (original magnification, 180). (C) Whole mount 3% O2-treated explant showing glomeruli labeled with PNA lectin (red) and FITC-labeled immunoreactive ACE (green/yellow) staining endothelial cells next and within glomeruli (original magnification, 1100). (E) 20% O2 explant showing no detectable Flt-1 immunoreactivity (original magnification, 180). (E) 20% O2 explant showing no Flk-1 immunostaining (original magnification, 180). (F) Whole mount 20% O2 explant showing PNA-labeled glomeruli (red) and no ACE immunostaining (original magnification, 180).

Article Snippet: Cultured explants were fixed in 4% paraformaldehyde, incubated with 0.1 U/ml neuraminidase for 60 min at 377C, washed, and incubated with rhodamine-labeled peanut lectin (PNA, Vector, Burlingame, CA) [7.5 mg/ml] and antiACE polyclonal antibody (1:200) for 90 min at 377C.

Techniques: Labeling, Staining, Immunostaining

Journal: Data in Brief

Article Title: Imaging data on characterization of retinal autofluorescent lesions in a mouse model of juvenile neuronal ceroid lipofuscinosis (CLN3 disease)

doi: 10.1016/j.dib.2020.106076

Figure Lengend Snippet: Autofluorescent lesions in the homozygous Cln3 Δex7/8 mouse retinal OPL/INL were negative for the cone photoreceptor marker peanut agglutinin. Peanut agglutinin (PNA), a lectin protein derived from the fruits of Arachis hypogaea and with high affinity for galactose-galactosamine disaccharide residues, stains cone photoreceptors [ , ]. Hypopigmented homozygous Cln3 Δex7/8 mice (10-17 month old) on an outbred 129Sv/Ev/CD1 mixed background, which is known to harbor additional CLN3-independent retinal degenerative genetic loci, were shown to have significantly decreased cone photoreceptors by PNA chromogenic IHC [ , ]. However, we did not observe overt photoreceptor loss by Cy5-PNA fluorescence in a homozygous Cln3 Δex7/8 mouse (22 month old) on a C57BL/6J background. In addition, autofluorescent lesions in the homozygous Cln3 Δex7/8 mouse retinal OPL/INL were negative for Cy5-PNA. Autofluorescence was collected with a 487.9 nm laser and a 525/25 nm emission filter. PNA IF was performed on retinal cyro-sections stained with Cy5-conjugated PNA (Vector CL-1075a) and confocal fluorescent images were acquired with a 638.6 nm laser and a 700/75 nm emission filter. (A) Representative DIC and confocal fluorescent images of homozygous Cln3 Δex7/8 mouse retina show autofluorescent lesions in the OPL/INL (green channel; pointed by yellow arrows) and Cy5-PNA fluorescence (red; pseudo-color) at the cone IS and OS and cone pedicles in the OPL/INL. Note that DIC images clearly show distinctive morphologies of RPE, OS, IS, ONL, OPL, INL, IPL, and autofluorescent lesions. (B) Representative DIC and confocal fluorescent images of WT mouse retina show autofluorescence (green) and Cy5-PNA fluorescence (red; pseudo-color). (C) Control fluorescent images of homozygous Cln3 Δex7/8 mouse retina acquired and processed exactly as in (A) except that Cy5-PNA staining was omitted. Note that bringing up signals in the pseudo-red channel revealed autofluorescent lesions rather than the typical Cy5-PNA staining patterns (A-B) in the OPL/INL.

Article Snippet: PNA IF was performed on retinal cyro-sections stained with Cy5-conjugated PNA (Vector CL-1075a) and confocal fluorescent images were acquired with a 638.6 nm laser and a 700/75 nm emission filter. (A) Representative DIC and confocal fluorescent images of homozygous Cln3 Δex7/8 mouse retina show autofluorescent lesions in the OPL/INL (green channel; pointed by yellow arrows) and Cy5-PNA fluorescence (red; pseudo-color) at the cone IS and OS and cone pedicles in the OPL/INL.

Techniques: Marker, Derivative Assay, Fluorescence, Staining, Plasmid Preparation