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Fig. 5 | Cell Communication and Signaling

Fig. 5

From: The multi-site docking protein Grb2-associated binder 1 (Gab1) enhances interleukin-6-induced MAPK-pathway activation in an SHP2-, Grb2-, and time-dependent manner

Fig. 5

Tyrosine 759 in gp130 is crucial for IL-6-induced Gab1 translocation and phosphorylation (a) HEK293 Gab1-KO cells were seeded on poly-L-lysine-coated glass cover slips and co-transfected with expression vectors for Gab1-WT-GFP and either EG (YYYYYY) or EG (YFYYYY). On the following day, cells were serum-starved for 4 h, subsequently placed into the incubation chamber of a laser scanning microscope and left for 30 min. Cells were treated with Epo (7 U/ml) to induce IL-6 signalling. Imaging was performed before and after treatment with Epo for up to 30 min as indicated. b HEK293 Gab1-KO cells were seeded and co-transfected with expression vectors for Gab1-WT-GFP and either EG (YYYYYY) or EG (YFYYYY). After 24 h, cells were serum-starved for 4 h. Cells were left untreated or treated with Epo (7 U/ml) for 15 min. Subsequently, cell lysates were prepared and proteins separated by SDS-PAGE. After Western blotting, membranes were stained for (p)STAT3, STAT3, (p)SHP2, SHP2, (p)Y627-Gab1, (p)S552-Gab1, Gab1, (p)Erk1/2, and Erk1/2. After stripping, membranes were stained for GFP. Representative results of n = 3 independent experiments are shown

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