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. Author manuscript; available in PMC: 2014 Feb 13.
Published in final edited form as: Oncogene. 2013 Jan 21;33(4):532–535. doi: 10.1038/onc.2012.619

Figure 1.

Figure 1

Oncogenic Ras requires activation for enhanced activity. (a) Lysates from HEK293 cells transfected with K-, N- and H-Ras cDNA were used to determine the specificity of corresponding antibodies by western blotting. (b) Active K-, N- and H-Ras were measured by a Raf pull-down assay from pancreas lysates of 2-month-old control and Panc-Ras mice (lanes a). For comparison with total corresponding Ras isoform proteins, 5% of total lysates used for pull-down assay were loaded to adjacent wells (lanes t) and probed with isoform-specific antibodies. Densitometry was performed using ImageJ software. (c) Active K-Ras was measured in isolated pancreatic acinar cells from control and Panc-Ras mice after EGF (50 ng/ml) stimulation. (d) Oncogenic Ras is partially active in cancer cells. Active K-, N- and H-Ras (lanes a) were compared with 10% of the corresponding Ras isoform levels used for pull-down assay (lanes t) in human pancreatic cancer cells (Bxpc3, Panc1 and Mpanc96) and pancreatic cancer cells derived from Panc-Ras mouse (K8484). Note all cells except BxPC-3 possess oncogenic K-Ras mutations.