Effects of GNF-2 and Dasatinib on Rat-1 cells expressing BCR/ABL-T315I. (A) Transformation assays measuring colony formation in soft agar. Rat-1 cells were retrovirally transduced with the BCR/ABL-T315I construct and seeded at 104 cells/well on soft agar in six-well plates in the presence of 2.5 or 5 μM GNF-2 and 300 nM Dasatinib. After 15 days, the colonies were counted. The means +/- SD of triplicates of two representative experiments are presented. (B) Focus formation assays. A total of 4x104 infected Rat-1 cells were plated per well in 24-well plates, grown for 72 h to confluence and incubated for 12 additional days in the presence of 2.5 or 5 μM GNF-2 and 300 nM Dasatinib. The plates were then washed, dried and stained with crystal violet. One representative of each of the two experiments performed in triplicate is given (34X magnification).