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Fig. 4. Molecular mechanism of Dkk2 gene function.
(A,B) At P10, Pax6 protein is detected in the nuclei of the
epithelial basal cells of the wild-type cornea (A) but is absent or excluded
from the nuclei of the mutant cornea (B). (C-G) X-gal staining on
paraffin sections taken from wild-type and
Dkk2–/– corneas of mice carrying a
ß-galactosidase (lacZ) gene under the control of
ß-catenin-responsive elements. At P2, significant lacZ
expression is detected in the limbus of the mutant cornea (D), but not in the
wild-type cornea (C). (E) Higher magnification view of D, showing localization
of the X-gal staining in the stroma of the limbus. The limbal
epithelial-stromal border is indicated. At P15, lacZ expression is
absent in the central region of a wild-type cornea (F), but detected in a few
hair follicles located in the central cornea of a mutant eye (G).
(H,I) RT-PCR analysis. Lef1 mRNA expression is
significantly upregulated in mutant corneas compared with wild-type corneas;
Shh mRNA expression is induced in mutant corneas and absent in
wild-type corneas. (J-M) Immunostaining and (N,O) X-gal
staining on paraffin-embedded sections from E15.5 corneas. K12 expression is
present (J) in wild-type cornea but absent (K) in a
Dkk2–/– cornea. Pax6 expression is present in
a wild-type cornea (L), but absent in a mutant cornea (M). TOPGAL expression
is induced in the limbal stroma region of a
Dkk2–/– cornea (O), but not in wild-type
cornea (N). Scale bars: in A, 50 µm for A,B; in C, 100 µm for C,D; in F,
100 µm for F,G; in J, 50 µm for J-M; in N, 150 µm for N,O.
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