Fig. 1.

GC RAS response to proliferative stimuli. a RAS is not activated at the GC by proliferative stimuli. MCF-7 cells transfected with HA-tagged KDELr HRAS (0.5 μg), unstimulated (0) or treated with EGF (50 ng/ml) or HRG (30 ng/ml) for the indicated times. Cells transfected with RASGRP1 (1 μg) (+) serve as positive control. GTP loading was assayed by GST-RBD pull-down (RAS-GTP-PD). b RAS activation by EGF is restricted to the plasma-membrane. MCF-7 cells transfected with constructs expressing cherry-HRAS and the RAS-GTP biosensor E3-R3 (RAF-RBD) (1 μg each) and stimulated for the indicated times. Scale bar = 10 μm