Fig. 5.

GISP prolongs EGF-induced activation of the ERK1/2. (A) HEK293 cells were transfected with either GISP or GISPΔ8-Mut vector. 48 h post-transfection, the cells were serum-starved for at least 6 h and then incubated with 50 ng/ml EGF for the times indicated. Samples were then washed in ice-cold PBS, scraped directly into loading buffer and then separated by SDS-PAGE. Blots were probed with anti-pERK1/2 antibody and also probed with anti-β-actin to ensure equal loading. These are the same samples described and used in Fig. 4. (B) Quantitative representation of the pERK1/2 western blot in (A) determined by image J analysis. Mean ± SEM from 4 independent experiments. *p < 0.05.