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Fig. 2. Localization of Wnt proteins in the chick neural tube. Wnt1 and GFP
(A-C) or Wnt3a and GFP (D-F) were expressed in the neural tube
following electroporation. Diagram to left depicts GFP (green) expression in
an electroporated neural tube; the boxed region is shown in A-F. GFP-positive
embryos were harvested 24 hours post-electroporation (HH stage 18/19), fixed
and cryosectioned prior to immunostaining with anti-Wnt1/Wnt3a antibodies.
Goat anti-mouse-Cy3 secondary antibodies were used to visualize Wnt proteins
(red). All images were collected by confocal microscopy. White arrows point to
perinuclear staining; arrowheads indicate immunopositive punctae. No punctae
were observed when primary antibody was omitted (data not shown). (G-O)
A wild-type chick embryo (HH stage 18) was cryosectioned and stained with Wnt1
primary antibody and a Cy3-labeled secondary antibody (red). Diagram to left
shows endogenous Wnt1 expression (red) in the neural tube; the boxed region is
shown in G-O. G and H are adjacent sections, as are I-L. In H, primary
antibody was omitted. In I, excess GST-Wnt1 blocking peptide was added. In
J-L, we compare the localization of Wnt1 protein (J) with that of
Wnt1 transcript (K). In M-O, we compare the localization of Wnt1
(red) with NCAM (green) in a single optical confocal layer. In M and O, white
arrows point to staining on or just interior to the plasma membrane, whereas
the white arrowheads indicate staining that is inside the cell. nt, neural
tube; no, notochord.
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