Fig. 5

KRAS-Mediated Intestinal Cell Proliferation following Loss of APC Requires β-Catenin

(A) Zebrafish embryos were injected with β-catenin^S45A mRNA (left) along with KRAS mRNA (middle). Also shown is a representative image of the APC-KRAS embryo (right). At 72 hpf, the embryos were fixed and photographed.

(B) The percent cyclops was analyzed (^*p < 0.01 WT versus β-catenin-KRAS, ^**p < 0.01 WT versus APC-KRAS).

(C) Protein was harvested from apc^mcr embryos treated with DMSO or NS-398 and subjected to western blot analysis for β-catenin (top) or vinculin (bottom).

(D) Wild-type uninjected or apc^mcr embryos injected with KRAS mRNA treated with VEH (top) or NS-398 (bottom) were stained by H&E (WT, left; APC-KRAS, right) and for DNA (magenta), PCNA (green), and β-catenin (green). TOPGFP-APC^mo-KRAS embryos were stained for GFP expression (purple). Boxes indicate the intestine.

(E) SW-480 cells treated with DMSO or NS-398 were stained for DNA (magenta) and β-catenin (green). Protein lysates were subjected to western blot analysis for β-catenin (top) and vinculin (bottom).

(F) Cells were subjected to MTT analysis (^*p < 0.05 versus DMSO; error bars, SEM).

Overlapping expression is shown in white. All images were captured using the same exposure and represent at least three independent experiments. Scale bar, 10 μm.