Fig. 5.

Reporter dynamics after exposure of HeLa cells to EGF and different signaling inhibitors. A: time course of the relative translocation response of the mKate2-ERK2 reporter (left) and the FoxO1-clover reporter (right) in cells incubated in SFM and then exposed to SFM or EGF (2.1 nM) ± different inhibitors for 90 min. B: time course of the relative translocation response of the mKate2-ERK2 reporter (left) and the FoxO1-clover reporter (right) in cells incubated in SFM and then exposed to SFM or EGF (2.1 nM) ± different inhibitors for 90 min. For A and B, population averages are presented (n = ~140 cells/incubation for each treatment group). For all experiments shown, the relative responsiveness of each reporter protein in each cell was normalized to the values at the start of imaging during incubation in SFM and scaled to the average peak EGF response. C: bar graph showing the average relative change in fluorescence intensity from an antibody array using HeLa cell lysates exposed to SFM or EGF (2.1 nM) ± different inhibitors for 15 min. Results are presented as means ± SE (n = 5 independent experiments). Phosphorylation sites are as follows: S6 ribosomal protein^Ser235/236, PRAS40^Thr246, p70 S6 kinase^{Thr421/Ser424}, Rsk1^Ser380, and ERK^{Thr202/Tyr204}.