Fig 4.

Systemic metastasis of cancer cells in nude mouse model. EGFP labeled breast cancer cell line MB231 and melanoma cell line C8161 were transfected with vectors expressing control shRNA or filamin-A targeting shRNA. Stable clones with filamin-A expression or filamin-A knock-down were isolated. Log-phase cells were injected into left ventricles of nude mice and body weight of injected mice were monitored weekly as shown in panel A. Four and eight weeks after injection, the mice inoculated with MB231-Con and C8161-Con displayed significant body weight loss (*p<0.05). Live images with EGFP marker were taken from anesthetic mice 8 weeks after injection for MB231 cells and 4 weeks for C8161 cells as shown in panel B. Hot spots of suspect tumor metastases were labeled with arrows and arrow heads for mouse injected with MB231-con and C8161-con cells respectively. Panel C shows the representative images of 94 μm sections proximal to distal physis. Panel D shows the quantitative analysis of physis wall thickness, which reduced significantly in mice with MB231-con tumor cells inoculation than in those injected with MB231-KD cells (p=0.022).