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

Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTB IgA MAbs block ricin attachment to ASF. Biotinylated ricin (8 ng/ml) was mixed with each of the indicated MAbs at a saturating concentration (10 μg/ml) and overlaid in triplicate onto ELISA plates coated with ASF. The plates were then washed and developed using avidin-HRP and TMB, as described in Materials and Methods. One hundred percent binding was defined as the value obtained for wells probed with ricin in the absence of antibody. Each bar represents the average, with SE, of two samples performed in duplicate.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Blocking Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Avidin-Biotin Assay, Binding Assay

Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTB IgA MAbs inhibit ricin binding to the luminal surfaces of the human duodenum. Biotinylated ricin (5 μg/ml) was incubated with each of the indicated IgA MAbs (20 μg/ml) and then overlaid for 1 h onto deparaffinized tissue sections of human duodenum. The sections were washed, labeled with streptavidin-FITC, and visualized by fluorescence microscopy. (A) Ricin uniformly labeled the luminal surfaces of human intestinal villi. Occasional goblet cells were also labeled (bottom center). (B) 33G2 reduced ricin binding to luminal surfaces to undetectable levels. (C) 35H6 reduced toxin binding to luminal surfaces, but residual staining was observed on the surrounding glass surfaces. Neither 23D7 (D) nor TEPC-15 (E) had an effect on ricin attachment to tissue sections compared to sections treated with ricin only. (F) Tissue section not labeled with ricin. Bar, 20 μm.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Binding Assay, Incubation, Labeling, Fluorescence, Microscopy, Staining

Journal:

Article Title: Immunoglobulin A Antibodies against Ricin A and B Subunits Protect Epithelial Cells from Ricin Intoxication

doi: 10.1128/IAI.02088-05

Figure Lengend Snippet: Anti-RTA IgA MAbs reduce transcytosis of ricin across polarized epithelial cell monolayers. Biotinylated ricin (200 ng) was incubated with the indicated IgA MAbs (2 μg in DMEM) in a volume of 0.2 ml for 1 h and then applied to the apical surfaces of polarized MDCK II cell monolayers. Eighteen hours later, ricin in the basolateral compartments was collected using avidin-agarose beads, which were then subjected to SDS-PAGE and Western blot analysis, as described in Materials and Methods. As positive and negative controls, samples of DMEM containing ricin (200 ng) (lane labeled “standard”) or no ricin (negative control) were treated in parallel.

Article Snippet: Unlabeled and biotinylated derivatives of ricin ( Ricinus communis agglutinin II), as well as polyclonal goat anti-RCA-I/II antiserum, were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Incubation, Avidin-Biotin Assay, SDS Page, Western Blot, Labeling, Negative Control