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- _4 value "To test our hypothesis that the Wnt/beta -catenin pathway is chronically activated in Foxl1 mutant mice we used indirect immunofluorescence to determine nuclear localization of beta -catenin as an indicator of the activated state of the Wnt pathway. Overall beta -catenin levels observed at low magnification, which mainly reflect beta -catenin contained in the adherens junctions, were comparable in the stomach of wild type and Foxl1 mutant mice (Fig. 2, A and C). However, at higher magnification we found a dramatic change in the subcellular localization of beta -catenin (Fig. 2, B and D). Foxl1 mutant mice had a larger number of nuclei with beta -catenin staining, indicating that the Wnt/beta -catenin pathway had indeed been activated in these cells. A similar increase in nuclear beta -catenin staining was also seen in the jejunum of Foxl1 mutant mice, confirming the correlation between increased proliferation and increased nuclear localization of beta -catenin (data not shown). To quantify the observed increase in nuclear beta -catenin, we performed Western blot analysis on cytoplasmic and nuclear extracts from Foxl1 mutant mice and their littermate controls (Fig. 3A). We found a significant increase in nuclear beta -catenin in the stomach and jejunum (2- and 2.3-fold, respectively) of Foxl1 mutant mice (Fig. 3B) confirming the immunofluorescence results, while cytoplasmic beta -catenin was unchanged. " provenance.
- _4 wasQuotedFrom 11555641 provenance.