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. 2008 Jul 18;105(30):10507–10512. doi: 10.1073/pnas.0711062105

Fig. 2.

Fig. 2.

eNOS increases the activation of Ras/ERK in T cells forming conjugates with antigen-pulsed APC. (A and B) GFP, eNOS-, and G2A-GFP cells were mixed with paraformaldehyde-fixed SEB-pulsed Raji cells for the times indicated. (A) Time course activation of ERK. (B) Time course activation of Ras determined by pull-down binding to GST-RBD-Raf-1 and subsequent immunoblotting with rabbit anti-pan-Ras mAb. GST-RBD-Raf-1 loadings are also shown. (C) Time course activation of Ras in eNOS-GFP cells pretreated with l-NAME or vehicle for 30 min before stimulation with SEB-pulsed Raji APC. (D) Time course activation of Ras in T cells conjugated with HA-pulsed HOM-2 cells. (E) RT-PCR of eNOS in T lymphoblasts from wild-type (WT) and eNOS-deficient (KO) mice. Control GAPDH mRNA expression is shown. (F) Ras activation in WT and eNOS KO mouse T lymphoblasts stimulated with CD3 Ab. As a control, eNOS KO cells were treated with 100 μM DETA-NO. Ras activation in C, D, and F was determined as in B. Each panel is representative of three independent experiments.