Increased invasiveness and tumorigenicity of MCF7 stable cell lines over-expressing GIG47. (A) MCF7 cells were transiently transfected with GIG47/pcDNA3.1 construct, and the cell pellet and culture media were separated by centrifugation. The cell pellet was lysed by a lysis buffer while culture media was concentrated. The GIG47 protein expression was analyzed by Western blot analysis with anti-V5 antibodies. Lane 1 and 2 are the cell lysate and concentrated culture media, respectively. (B) Western blot analysis of GIG47 protein levels in established MCF7 stable cell lines. (C) Effect of GIG47 over-expression in in vitro invasion of MCF7 cells. Invasion analysis of three clones was performed in serum-rich medium (50% FBS). Experiments were repeated at least three times. MDA-MB-231 cells were used as a positive control. (D) Increased tumorigenicity of GIG47 over-expressing MCF7 cells. MCF7-GIG47 clones (G3 and G7) and MCF7-pcDNA3.1 cells were inoculated into mammary fat pads of athymic nude mice. The tumor growth was measured every week after the inoculation. (E) The cell growth rate of NIH/3 T3 overexpressing GIG47 was significantly increased compared to control cells. (F) GIG47-overexpressing NIH/3 T3 clones exhibited the increased tumor growth rate compared with NIH/3 T3 cells vector control.