Journal: eLife

Article Title: An Eya1-Notch axis specifies bipotential epibranchial differentiation in mammalian craniofacial morphogenesis

doi: 10.7554/eLife.30126

Figure Lengend Snippet:

Article Snippet: antibody , Rat monoclonal anti-Jagged1 , DSHB (Iowa, USA) , Ts1.15h, RRID: AB_528317 , 1/300, IHC.

Techniques: Recombinant, Transfection, Protease Inhibitor, In Situ, Sequencing, Cloning, In Situ Hybridization, Construct, Generated, Plasmid Preparation, Mutagenesis, Software

Journal: Endocrinology

Article Title: Disruptions of global and JAGGED1-mediated notch signaling affect thyroid morphogenesis in the zebrafish.

doi: 10.1210/en.2011-1888

Figure Lengend Snippet: FIG. 8. Model describing the role of Notch signaling and Jag1-Notch interaction in zebrafish thyroid development. The thyroid primordium originates from the pharyngeal endoderm by 24 hpf. Our data indicate that Notch signaling originating from the endodermal layer itself or from the surrounding developing tissues plays a fundamental role in limiting the number of thyroidal differentiating precursors (double dotted arrow, 1, left side of the panel). The combined expression of nkx2.1a, pax2a, hhex, and pax8 is distinctive of the thyroid follicular cells. jag1b is mainly expressed starting from at 48 hpf; nevertheless, we can not completely exclude jag1b contribution in earlier phases of thyroid development (“jag1b?”), due to its spotted pattern of expression in the region of the thyroid primordium origin at 24 hpf; jag1a is mainly expressed at 72 hpf, in the most anterior portion of the first thyroid follicle. By 72 hpf, the thyroid tissue starts to elongate posteriorly and produce T4. Jag1-Notch signaling sustains these steps and the maintenance of thyroid cells (dotted black arrow, 2). jag1 loss-of- function conditions impair thyroid development and function (black arrow, 3). T, Thyroid; ch, ceratohyal; h, heart. Stages, upper part of the panel; 24 and 36 hpf, transversal views; from 48 to 120 hpf, ventral views.

Article Snippet: For Jag1 immunofluorescence on mouse thyroid tissue, sections (5 m) cut from OCT-embedded tissue blocks were blocked with 1% BSA/2% donkey serum and immunostained overnight at 4 C with Jagged1 polyclonal antibody (1:100, H-114; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Expressing

Journal: Brain

Article Title: Notch-1 signalling is activated in brain arteriovenous malformations in humans

doi: 10.1093/brain/awp246

Figure Lengend Snippet: Immunohistochemical detection of Notch-1 ligands and Hes-1 in human brain AVMs. (A) Dll4, Jagged-1 and Hes-1 expression (black) was increased in human brain AVMs (left panels, low-magnification; middle panels, high-magnification) compared with normal human middle cerebral artery (right panels). A few non-vascular cells in normal control brain also expressed Hes-1 (arrow). (B) Notch1, Jagged-1, Dll4, NICD and Hes-1 were expressed or activated in vascular cells of E17 mouse brain. (C) Immunostaining was performed on H9 human embryonic stem cells using (left to right) anti-Jagged-1 (green) and anti-NICD (red); anti-Notch-1 (green); anti-Dll4 (green) and anti-Hes1 (green). DAPI (blue) was used to counterstain nuclei.

Article Snippet: Recombinant human Notch-1/Fc chimera, recombinant human Jagged-1/Fc chimera, recombinant human Dll4 and Hes-1 protein (all from R&D System) were used as positive controls.

Techniques: Immunohistochemical staining, Expressing, Immunostaining

Journal: Brain

Article Title: Notch-1 signalling is activated in brain arteriovenous malformations in humans

doi: 10.1093/brain/awp246

Figure Lengend Snippet: Western blot analysis and cell-type localization of Notch-1 ligands in human brain AVMs. (A) Protein was isolated from the sources listed in the legend to Figure 1, and from normal mouse cerebral cortex (M-CTX). Western blotting was performed using antibodies against the Notch-1 ligands Dll4 (top panel) and Jagged-1 (middle panel), with anti-actin to control for differences in protein loading (bottom panel). (B) Recombinant human Notch-1, Jagged-1, Dll4 and Hes1 were loaded on gels and probed with the indicated antibodies, showing that the antibodies used recognize authentic antigens. Each pair of gel lanes was loaded with 100 ng (lane 1) or 500 ng (lane 2) of total protein. (C) Double-label immunostaining of human brain AVM sections shows that αSMC-expressing vascular smooth muscle cells also express Jagged-1 (top panel) and Dll4 (bottom panel). DAPI (blue) was used to counterstain nuclei. (D) Triple-label immunostaining of human brain AVM shows co-expression of Jagged-1 (red) and Hes1 (green) in CD31-immunopositive (purple) endothelial cells. DAPI (blue) was used to counterstain nuclei.

Article Snippet: Recombinant human Notch-1/Fc chimera, recombinant human Jagged-1/Fc chimera, recombinant human Dll4 and Hes-1 protein (all from R&D System) were used as positive controls.

Techniques: Western Blot, Isolation, Recombinant, Immunostaining, Expressing