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. 1999 Jun;10(6):1799–1809. doi: 10.1091/mbc.10.6.1799

Figure 3.

Figure 3

Activated but not wild-type or dominant-negative R-Ras can reverse the suppressive effect of activated H-Ras(G12V). Top, αβ-py cells were cotransfected with an expression vector encoding H-Ras(G12V). They were simultaneously cotransfected with 2 and 4 μg of a plasmid encoding wild-type R-Ras (●) or R-Ras(S43N) (⋄) and 0.5, 1, 2, and 4 μg of a plasmid encoding R-Ras(G38V) (▪). After 48 h, integrin activation was determined by PAC1 binding. Depicted is the activation index in the presence of H-Ras(G12V) ± SE of three independent determinations. Bottom, The immunoblot analysis of cell lysates illustrates the expression of wild-type (Wt) R-Ras and R-Ras(G38V) in the presence of H-Ras(G12V). Twenty micrograms of cell lysate from each transfection were separated on a 4–20% gradient gel and immunoblotted with either an anti-HA antibody, 12CA5, or the anti-Myc antibody 9E10. The R-Ras(S43N) was also well expressed, as determined by immunoblot analysis of lysates from R-Ras(S43N)–transfected cells (our unpublished observations).