Figure 3

Time-course and PKC-dependency of ERK and Akt phosphorylation in HCT116 cells. A, Time-course of neurotensin-induced phosphorylation of ERK and Akt. Quiescent cells were stimulated with 1 μM neurotensin (NT) for the times indicated. Cells were harvested for subsequent Western analysis as described in Methods. Results represent one typical of three independent experiments. B, Effect of increasing concentrations of neurotensin on phosphorylation of ERK and Akt. Quiescent cells were stimulated with increasing concentrations of neurotensin for five minutes. Cells were harvested for subsequent Western analysis as described in Methods. Results represent one typical of three experiments. C, Role of PKC in NT-induced phosphorylation of ERK and Akt. Quiescent cells were pretreated with vehicle (0.9% NaCl) or 3.5 μM GF109203X for 30 min, before stimulation with 1 μM NT, 1 μM TPA, or 5 nM EGF for five min and then harvested for subsequent Western analysis. Results represent one typical of eight independent experiments.