A, immunofluorescence shows that untreated G361 Wnt5Alow cells stain strongly for E-cadherin (green staining) but have no vimentin expression (red staining). B, by 12 h of rWnt5A (0.1 μg/ml) treatment, E-cadherin (green staining) is lost from the membrane and cytoplasm in the untreated cells, and vimentin is significantly up-regulated. C, E-cadherin (green staining) is abundant and Snail (red staining) is minimal in untreated G361 cells. D, treatment with rWnt5A for 12 h results in an increase in Snail expression and a decrease in E-cadherin. E, negative control cells incubated with secondary antibodies only.