Transfection of a PKCδ construct inhibits SCF-stimulated growth of IC2 cells expressing WT Kit, but enhances factor-independent growth of cells expressing D814Y mutant Kit. IC2 cells expressing WT Kit or the D814Y mutant were transfected with constructs coding for wild-type or dominant-negative PKCδ by electroporation. Control cells were electroporated with empty vector (MTH). (A) WT Kit transfectants were cultured for 48 hours in RPMI media supplemented with 10% FBS and 100 ng/mL murine SCF. (B) D814Y Kit transfectants were cultured for 48 hours in RPMI media supplemented with 10% FBS alone or (C) in RPMI media supplemented with 10% FBS (▦) and 100 ng/mL murine SCF (▧). Cells were pulsed with 3H-thymidine and harvested after a 6-hour incubation period. Mean counts of quadruplicate samples are shown above. These data are representative of 5 independent trials. Error bars indicate standard deviation.
