Effect of Fmn2 protein expression on anchorage of NIH/3T3 cells. Cells were transiently transfected with pCMV empty vector, with the pCMV-Fmn2 vector, and the Ras (EJ 6.6) expression vector. Transfected or control cells (104) were plated in soft agar as described in “Colony formation in soft agar.” After 4 weeks, the number of colonies (at least twice larger than colonies from controls) was scored (A). The results represent the average of 3 independent experiments. Statistical analysis was performed using one-way analysis of variance (P < .05). Cells were observed with an optical microscope (Ernst Leitz, 6MBH Wetzlar), and representative fields were photographed using a numerical camera (Nikon coolpix 4500; original magnification ×40). Images were analyzed using NIH ImageJ software Version 1.42l. Cells were left untransfected (B) or transfected with Ras EJ 6.6 (C), empty vector (D), or pCMV-Fmn2 (E).
