Fig. 5

MAPK and PI3K signalling are crucial for Gab1 plasma membrane recruitment in MDA-MB-468 cells. A MDA-MB-468 cells were seeded on poly-L-lysine-coated glass cover slips and cultivated in phenol red-free RPMI 1640. After 24 h, cells were transfected with an expression vector for either murine Gab1-GFP, murine Gab1-S552A-GFP or murine Gab1-ΔPH-GFP. On the following day, cells were placed into the incubation chamber of a laser scanning microscope. Imaging was performed after 30 min equilibration. Representative results of n = 3 independent experiments are shown. B MDA-MB-468 cells were seeded on poly-L-lysine-coated glass cover slips and cultivated in phenol red-free RPMI 1640. After 24 h, cells were transfected with an expression vector for murine Gab1-GFP. On the following day, cells were placed into the incubation chamber of a laser scanning microscope and left for 30 min. Cells were treated with DMSO, U0126 (10 µM) and/or LY294002 (40 µM) for 30 min. Imaging was performed before and after treatment. Representative results of n = 3 independent experiments are shown