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. 2015 Jun 24;6:7354. doi: 10.1038/ncomms8354

Figure 4. Dimeric Grb2 inhibits while monomeric Grb2 promotes MAP kinase activity.

Figure 4

(a) Stable HEK293T cells overexpressing FGFR2–GFP (as control, C) and strep-tagged wild-type Grb2 or the dimerization-defective Y160E mutant were stimulated with 50 ng/ml FGF2 or EGF for 15 and 5 min, respectively. Cell-lysates were analysed for phospho-ERK (pERK), total ERK and Grb2 expression levels using specific antibody and Odyssey infra-red imaging. (b) HEK293T cells overexpressing FGFR2–GFP with indicated strep-tagged Grb2 in serum were lysed and subjected to strep-tactin affinity purification. The resulting co-precipitated complexes along with input cell-lysates were analysed for SOS and FRS2 binding using respective antibody. The immunoblot was also re-probed for Grb2 as a loading control. The data presented are representative of three independent experiments.