Fig. 2.

GC RAS response to apoptogenic stimuli. a Apoptogenic stimuli activate RAS at the GC. MCF-7 cells transfected with HA-tagged KDELr HRAS (0.5 μg), unstimulated (0) or treated with TGF-β (5 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 in endomembranes induced by TGF-β. MCF-7 cells transfected with constructs expressing cerulean-HRAS and the RAS-GTP biosensor E3-R3 (RAF-RBD) (1 μg each) and stimulated for the indicated times. GC was revealed by the RFP Golgi probe. Insets show areas of prominent RAS-GTP accumulation. Scale bar = 10 μm. See also Supplementary Fig. 1