Immunodetection of Neuronal Markers

Immunodetection of neuronal markers was carried out as previously described [53 (link), 54 (link)]. Primary antibodies used were: rabbit anti-doublecortin (Cell Signaling Technology Inc.), monoclonal anti-NeuN (Millipore), goat anti-FZD1 (R&D Systems), goat anti-FZD1 (LifeSpan Biosciences, Inc.), rabbit anti-SOX2 (Cell Signaling Technology Inc.), monoclonal anti-GFAP (Sigma-Aldrich), monoclonal anti-Nestin (Millipore), rabbit anti-Ki67 (Abcam). As secondary antibodies, Alexa (Molecular Probes) and DyLight (Abcam) conjugated antibodies were used. NucBlue (Life Technologies) was used as nuclear dye. Slices were mounted on gelatin-coated slides with Fluoromont-G (Electron Microscopy Sciences). Double-labeled sections were analyzed by confocal laser microscopy (Olympus FV 1000). Image analysis and z-projections were made with ImageJ software (NIH, USA).

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Mardones M.D., Andaur G.A., Varas-Godoy M., Henriquez J.F., Salech F., Behrens M.I., Couve A., Inestrosa N.C, & Varela-Nallar L. (2016). Frizzled-1 receptor regulates adult hippocampal neurogenesis. Molecular Brain, 9, 29.

Publication 2016

rabbit anti-doublecortin monoclonal anti-NeuN goat anti-FZD1 rabbit anti-SOX2 monoclonal anti-GFAP monoclonal anti-Nestin rabbit anti-Ki67 Alexa DyLight NucBlue Fluoromont-G FV 1000

Antibodies Doublecortin Electron microscopy Gelatin Gfap Goat Laser microscopy Molecular probes Nestin Neuronal Rabbit Sox2

Corresponding Organization :

Other organizations : Universidad Andrés Bello, Universidad de Los Andes, Chile, Hospital Clínico de la Universidad de Chile, University of Chile, Pontificia Universidad Católica de Chile

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Variable analysis

independent variables

Primary antibodies used were: rabbit anti-doublecortin (Cell Signaling Technology Inc.), monoclonal anti-NeuN (Millipore), goat anti-FZD1 (R&D Systems), goat anti-FZD1 (LifeSpan Biosciences, Inc.), rabbit anti-SOX2 (Cell Signaling Technology Inc.), monoclonal anti-GFAP (Sigma-Aldrich), monoclonal anti-Nestin (Millipore), rabbit anti-Ki67 (Abcam)

dependent variables

Immunodetection of neuronal markers

control variables

As secondary antibodies, Alexa (Molecular Probes) and DyLight (Abcam) conjugated antibodies were used
NucBlue (Life Technologies) was used as nuclear dye
Slices were mounted on gelatin-coated slides with Fluoromont-G (Electron Microscopy Sciences)

controls

No positive or negative controls were explicitly mentioned

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