Figure 2. Interfering with the expression of Merlin promotes metastasis in HCC cell lines.

(a) Lysates of HCCLM3 cells transfected with Merlin shRNAs were immunoblotted with the indicated antibodies. (b) Generation of shMerlin stable HCCLM3, Huh7 and L02 cell lines. (c) Transwell and (d) invasion assays were used to test the migration and invasion abilities of HCC cells. The respective infiltrated cell numbers are shown in the lower panes. (Scale bar, 50 μM). Results are shown as the mean±s.d., n=3. **P<0.01, ***P<0.001, based on the student's t-test. (e) Representative lung tissue sections of BALB/C mice killed at 13 weeks from each group are shown (H&E; Magnification : × 100, Scale bar, 300 μM ). The arrows indicate lung metastatic tumours (lower images). The average number of lung metastatic tumours per lung in each group (n=5) was calculated (upper panel). Results are shown as the mean±s.d., n=5. **P<0.01, based on the student's t-test. (f) Huh7 cells were treated in vitro with GTPγS or GDP to activate Rho and Cdc42. Cell lysates (50 μl at 1 mg ml⁻¹) in Western blotting showed active Rho and Cdc42 levels in the shMerlin cell line and the control.