Inhibition of PKCδ has different effects on growth of IC2 cells expressing WT and D814Y Kit. IC2 cells were cultured with the PKCδ selective inhibitor rottlerin or the conventional isoform PKC inhibitor Gö6976 in cell proliferation assays. (A) IC2 cells expressing WT or D814Y mutant Kit were incubated with the indicated concentrations of rottlerin for 1 hour 20 minutes and then cultured for 72 hours in RPMI media supplemented with 10% FBS and 100 ng/mL murine SCF (WT Kit; ♦) or media supplemented with 10% FBS alone (D814Y Kit; ▪). (B) IC2 cells expressing D814Y mutant Kit were incubated with the indicated concentrations of rottlerin for 1 hour 20 minutes and then cultured for 72 hours in RPMI media supplemented with 10% FBS and 100 ng/mL murine SCF (▪) or media supplemented with 10% FBS alone (♦). Cells were pulsed with 3H-thymidine and harvested after a 4-hour incubation period. Mean counts of quadruplicate samples exposed to the drug were normalized to the means of samples that were cultured without drugs. These data are representative of the results of 4 independent trials. (C) IC2 cells expressing WT or D814Y mutant Kit were incubated with the indicated concentrations of Gö6976 for 1 hour and then cultured for 72 hours in RPMI media supplemented with 10% FBS and 100 ng/mL murine SCF (WT Kit; ♦) or media supplemented with 10% FBS alone (D814Y Kit; ▪). (D) IC2 cells expressing D814Y mutant Kit were incubated with the indicated concentrations of Gö6976 for 1 hour and then cultured for 72 hours in RPMI media supplemented with 10% FBS and 100 ng/mL murine SCF (▪) or media supplemented with 10% FBS alone (♦). Cells were pulsed with 3H-thymidine and harvested after a 4-hour incubation period. Mean counts of quadruplicate samples exposed to the drugs were normalized to the means of samples that were cultured without drug. These data are representative of the results of 3 independent trials. Error bars indicate standard deviation.
