Figure 5: UVRAG^FS activates Rac-1 and promotes tumour metastasis in vitro and vivo.

(a) Rac-1 activation by UVRAG^FS. Western blot shows a pull-down experiment to detect GTP-bound Rac1 in SW480.UVRAG^FS cells and on drug treatment. Histogram shows quantification from three independent experiments. (b) Representative images of scratch-wound healing exhibit the motility of SW480.UVRAG^FS cells. Cell motility into the wound area was taken at 0 and 16 h as marked by red lines. Wound-healing index was quantified (right). Data are the means±s.d. (n=3). **P<0.01; ***P<0.001; NS, not significant. Histogram shows quantification from three independent experiments. (c,d) UVRAG^FS enhances tumour metastasis in mice inoculated by intrasplenic injection of SW480.Vector and SW480.UVRAG^FS cells. Schematic depiction of the procedure (c, left) and representative images (c, right) of upper abdominal organs at 8-week post-injection are shown (c). The number of metastatic nodules (liver and lung) was quantified (d). H&E staining was performed on serial sections of metastatic tumours (M) and normal (N) liver and lung are shown below. Scale bar, 1 mm. Arrows in c represent the metastasis foci. Results are representative of 10 mice per group. Fisher’s exact test was used. (e) Immunohistochemistry analysis of autophagy, apoptosis and EMT status of primary tumours (left panel) and metastasis nodules (right panel). The bar graphs represent the quantification of the indicated protein markers. Data are the means±s.d. (n=3). *P<0.05. Scale bar, 50 μm. (f) Western blot of the EMT-related protein expression in SW480 cells expressing UVRAG^FS and on Taxol treatment.