Fig. 3.

eNOS selectively activates N-Ras in antigen-stimulated T cells. (A) Time course activation of endogenous N- and K-Ras in eNOS- and G2A-GFP cells stimulated with CD3 Ab. Extracts of cells expressing HA-tagged N-, H-, and K-Ras were loaded as controls of antibody specificity. (B) eNOS- and G2A-GFP cells transiently expressing CFP-N-Ras or -K-Ras were stimulated with SEB-pulsed Raji APC for the times indicated. The expression of activated and total Ras was analyzed as in A. The loadings of GST-RBD-Raf-1 (RBD) are also shown. Each panel is representative of three independent experiments.