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Fig. 8. Upregulation of β-catenin after Wt1 deletion in
Sertoli cells. (A) Nuclear localization of β-catenin in
Sertoli cells of Wt1fx/-; AMH-Cretg/+ mice at
E15.5. β-Catenin protein was stained by an anti-β-catenin antibody
(green) and nuclei were counterstained with DAPI (blue). β-Catenin was
normally located on Sertoli cell membrane in control testes (left panel,
arrowheads). However, β-catenin was detected in Sertoli cell nuclei in
testes of Wt1 conditional knockout mice (right panel, arrows). Scale
bar: 20 µm. (B) Immunofluorescent images of isolated primary Sertoli
cells stained with WT1 (green). Nuclei were counterstained with DAPI (blue).
Over 90% of the cells were WT1 positive. (C) Real-time PCR
quantification of Wt1 expression in isolated primary Sertoli cells of
mutant and control mice, with or without 4OH-tamoxifen treatment. Numerical
data present mean±s.e.m. of relative expression of Wt1 in
three independent experiments. Column 1, Wt1fx/fx; +/+
with treatment; column 2, Wt1fx/-;
CAGGCre-ERtg/+ without treatment; column 3,
Wt1fx/-; CAGGCre-ERtg/+ with treatment.
Wt1 expression was significantly reduced upon 4OH-tamoxifen treatment
in Wt1fx/-; CAGGCre-ERtg/+ Sertoli cells.
(D) Western blot to analyze the expression levels of total
β-catenin and the active form of β-catenin in isolated primary
Sertoli cells of mutant and control mice, with or without 4OH-tamoxifen
treatment. The levels of total β-catenin and the active form of
β-catenin were upregulated upon 4OH-tamoxifen treatment in
Wt1fx/-; CAGGCre-ERtg/+ Sertoli cells
(quantification on the right).
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