FIG. 6.

(A) In vitro kinase assay demonstrating that the PIM3 K69M mutant has attenuated intrinsic kinase activity. The lysates used on the immunoblot represent an equal proportion of product immunoprecipitated from 293T cells transfected with either wild-type PIM3 or PIM3 K69M. The amounts of PIM3 protein present in the immunoprecipitated product are equal between the wild-type and K69M mutant samples. (B) Immunoblot analysis demonstrating expression of EWS/FLI1 and PIM3 K69M in the NIH 3T3 cell lines utilized in the tumorigenesis assay. (C) Growth curves of NIH 3T3 cells expressing PIM3 K69M or empty vector. The graph is representative of two independent experiments performed in quadruplicate.