Fig. 7. Charged amino acids at residue 551 mimic the ERK stimulus to induce translocation of Gab1 to the plasma membrane. HEK293T cells were transfected with expression vectors for Gab1-GFP wild-type proteins or mutants where Ser551 and Ser567 are replaced by alanine or glutamate (A), or mutants where only Ser551 was replaced by alanine or glutamate (B), or mutants where only Ser567 was replaced by alanine or gluatmate (C) together with vectors encoding EpoR-gp130 chimeric receptors. 24 hours after transfection, cells were transferred to poly-L-lysine-coated coverslips. The next day, cells were starved for 3 hours in KRH and transferred into the perfusion chamber of a laser-scanning microscope. After 20 minutes the cells were stimulated with Epo (7 U/ml). Gab1 localisation is shown before stimulation and after 20 minutes of Epo stimulation.