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Figure 3 | BMC Systems Biology

Figure 3

From: Spatio-temporal modeling of signaling protein recruitment to EGFR

Figure 3

Analysis and simulation of the reaction kinetics between the four adaptors and EGFR. (A-B) Membrane sheets were prepared from serum-starved, batimastat-treated A431 cells without (A) or with EGF stimulation (B). Sheets were labeled with 5 nm gold reagents recognizing Shc. Circles in (A, B) highlight Shc label on these membranes. Bars, 0.1 μm. (C-F) Quantitative values of Shc, Stat5, PLCγ1, and Grb2 immunogold labeling on 3 μm2 area of membrane, reported as an average of at least 10 membranes. Blots in C-F show results of fractionation experiments, where crude cytosol and membrane fractions were prepared, proteins separated by SDS-PAGE and membranes blotted for Shc, Stat5, PLCγ1 and Grb2. In (G-I), blots report co-precipitation of Shc, Stat5 and PLCγ1 with EGFR over a time course of EGF stimulation. Bands were quantified by densitometry and plotted as density of the bands. In (J-M), simulations of reaction kinetics between the four adaptors and EGFR using experiment-fitted values produce results (black solid line) similar to experimental data (grey dashed line).

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