Fig. 3.

(A) Exposure to Wnt3a causes an evident phenotypic transformation of mouse mammary epithelial cells into long chord-like bundles. (B–F) C57MG cells cultured in the presence of Wnt3a and compounds and/or DMSO stained with DAPI (blue) and FITC-conjugated phalloidin (green) for quantitative high-content image analysis. Candidate compounds inhibit (arrowhead in D–F) the acquisition of the chord-like phenotype seen in DMSO+Wnt3a-treated cells (arrow in C). The algorithm used to quantify this phenotypic transformation identifies intracellular actin fibers (blue overlay in B′–F′) and calculates the anisotropy of the actin fiber alignment within each individual cell (yellow mask outline in B′–F′). (B″–F″) Histogram plot of anisotropy of fiber alignment in cell populations (n ≥ 8,000) treated with individual candidate compounds shows normal distribution in controls; normal peak maxima is highlighted by the square bracket (B″). Treatment with Wnt3a+DMSO results in a skewed distribution of anisotropy (C″), which is rescued by treatment with candidate compounds (D″–F″). (Scale bar: 5 μm.) (G) Quantification of the β-cat target gene, WISP1, in response to treatment with candidate compounds. Error bars show range of variation from mean. (H) MCF7-S37Aβ-cat-HA cells show increased accumulation of Axn2 mRNA compared with control cells, which is inhibited by treatment with candidate compounds. (I) MCF7-S37Aβ-cat-HA cells exhibit invasive potential as assayed by the Boyden chamber invasion assay. Treatment with candidate compounds results in a marked decrease in the number of invading cells.