Effect of TIAM1 on Cell Migration and In Vivo Invasion

(A) Representative images of SW480 cells or SW620 cells migrating through Transwell inserts toward serum for 24 hr. Cells were transfected with siRNA for TIAM1 or TAZ/YAP either alone or in combination, as indicated. Scale bars, 150 μm. Graphs show average number of migrating cells normalized to negative control siRNA (siNT) and are presented as mean ± SEM (with siNT set as 100) (unpaired t test: ^∗p < 0.05, ^∗∗∗p < 0.001).

(B) Representative images of parental SW480 or the CRISPR-mediated TIAM1 knockout clones (TIAM1-KO #1 and # 2) migrating through Transwell inserts toward serum for 24 hr. Cells were transfected with NLS-TIAM1 or empty vector (EV) as control. Scale bars, 150 μm. Graph shows average number of migrating cells normalized to EV and are presented as mean ± SEM (EV transfected parental cells set as 100) (unpaired t test: ^∗p < 0.05, ^∗∗∗p < 0.001).

(C) Representative images at 4 days post injection of parental green fluorescent SW480 and SW480-TIAM1-KO#1 cells, non-induced (Minus Dox), or inducibly expressing (Plus Dox) either FL-TIAM1 or NLS-TIAM1 engrafted into the pericardial cavity of zebrafish embryos. Upper row shows epifluorescence of injected cells. White dotted line indicates the pericardial cavity. Lower row shows merge with bright field. Scale bar, 100 μm. Graphs show quantitation of invasion. Mean values ± SEM from three independent experiments (Kruskal-Wallis one-way ANOVA and Dunn's post hoc test: ^∗p < 0.05, ^∗∗p < 0.01).

See also See also Figure S7.