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. Author manuscript; available in PMC: 2011 May 28.
Published in final edited form as: Cell. 2010 May 20;141(5):884–896. doi: 10.1016/j.cell.2010.03.054

Figure 3. Nuclear ERK activation dynamics and the effect of dusp downregulation on c-fos mRNA duration.

Figure 3

A. Model predictions for nuclear ppERK time courses. Ligand concentrations are 10 nM (EGF, blue and HRG, red). B and C. Quantified nuclear ppERK dynamics based on cell images obtained from Duolink technology (EGF, blue and HRG, red; representative image is shown in Table S5A). Each data point is the average response based on ~180 individual cells in three independent experiments, and error bars correspond to standard error based on the three replicates. Solid lines denote in silico simulations, and dashed lines denote simulation standard deviation. For normalization, raw quantified data are divided by the 5 min. time point of each respective ligand dose. Shading corresponds to the nuclear ppERK profile between 15 and 60 min. D. Spatially-resolved ERK activation dynamics observed by immunofluorescence. Total ERK (green) is shown on the right and ppERK (red) is shown on the left. E,F,K,L. Measured vs. predicted effects of dusp downregulation on c-fos mRNA expression induced by 10 nM EGF (E and K) or HRG (F and L). Solid and dashed lines correspond to model simulations and their standard deviation, respectively. The dusp downregulation was modeled as an increase in the dusp mRNA degradation rate. Simulations in Panels E and F correspond to the initial model (Fig. 1E, black lines only) and simulations in Panels K and L correspond to the refined model (Fig. 1E, black and orange lines). G and H. c-fos mRNA expression in response to two 1 nM pulses of (G) EGF or (H) HRG. Arrows denote the second stimulation time. I and J. Effects of cycloheximide on c-fos mRNA expression induced by 10 nM EGF (I) or 10 nM HRG (J). For Panels E-L, error bars denote the standard error from three independent experiments. Note that Panels I and J have different y-axis scales. In Panels E, F, and I-L, data values are relative to their respective 30 min. HRG control point. Solid and dashed lines correspond to model simulations and their standard deviation, respectively. Simulations done with the refined model are indicated. See also Table S5.