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Fig. 4. Interaction of endogenous ß-catenin and Sox17. (A) A 160 µM
mid-sagittal optical z-series of anti-ß-catenin
immunofluorescence in a Xenopus midgastrula shows both membrane bound
and nuclear ß-catenin in the deep endodermal cells. The intense staining
throughout the animal cap and mesoderm is the result of the cells in this
region of the embryo being much smaller than in the endoderm (therefore the
optical stack is several cells thick in these regions, resulting in an almost
uniform staining). The right panel shows a higher magnification of the
endoderm (white box) with the stained nuclei clearly visible. Dorsal/anterior
towards the left. (B) Western blotting of whole cell, cytosolic and nuclear
extracts (10 µg of protein each) from human SW480 colorectal cancer cells
with anti-tubulin (cytosol antigen), anti-histone H1 (nuclear antigen),
anti-Sox17 and anti-ß-catenin antibodies. SW480 cells express both
endogenous Sox17 and ß-catenin in the nuclear fraction. (C) After
immunoprecipitation of the nuclear extract with either anti-Sox17 or anti-HA
(as a negative control) antibodies, associated ß-catenin protein was
detected by western blotting. Endogenous ß-catenin co-immunoprecipitated
with nuclear Sox17. The precipitation of Sox17/ß-catenin complexes can be
competed by the addition of Sox17 peptide recognized by the anti-Sox17
antibody but not by peptides to other regions of Sox17.
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