Fig. 3.

Reporter dynamics after exposure of HeLa cells to different concentrations of growth factors. A and B: time course of the relative translocation responses of the mKate2-ERK2 reporter (A) or the FoxO1-clover reporter (B) in cells incubated in SFM and then exposed to several concentrations of insulin-like growth factor-I (IGF-I) for 90 min. Population averages are presented (n = ~140 cells/incubation for each treatment group). For all experiments illustrated, the relative responsiveness of each reporter protein in each cell was normalized to values at the start of imaging during incubation in SFM and scaled to the average peak response. C: time course of the relative translocation response of the mKate2-ERK2 reporter (red traces) and the FoxO1-clover reporter (green) in individual cells incubated in SFM and then exposed to IGF-I (50 pM) for 90 min. D and E: time course of the relative translocation response of the mKate2-ERK2 reporter (D) and the FoxO1-clover reporter (E) in cells incubated in SFM and then exposed to different growth factors for 90 min. Population averages are presented (n = ~140 cells/incubation for each treatment group). For all experiments pictured, the relative responsiveness of each reporter protein in each cell was normalized to values at the start of imaging during incubation in SFM and scaled to the average peak response. F: dot plot of ERK and Akt activity, as indicated by responses of the mKate2-ERK2 reporter at 5 min and the FoxO1- clover reporter at 10 min in individual cells after incubation with SFM (black), transforming growth factor-α (TGF-α, 1.67 nM, purple), EGF (1.67 nM, blue), IGF-I (250 pM, green), insulin (1.0 nM, orange), or HGF (1.72 nM, red) (n = ~150 cells/treatment group).