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. 2011 Sep 19;6(9):e24931. doi: 10.1371/journal.pone.0024931

Figure 3. Active Ras can recapitulate CD28-mediated anergy prevention.

Figure 3

A. CAR Tg Th1 cells were left untransduced or transduced with empty vector or Ras61L. They were then rested or submitted to anergy-inducing conditions as described in the materials and methods. A. These cells were then restimulated overnight with non-coated or antibody coated beads or PMA and ionomycin and supernatants were assessed for IL-2 production by ELISA. Error bars represent the standard deviation of mean values from triplicate samples from the same experiment. Cells transduced with active Ras prior to anergy induction produced significantly more IL-2 in response to CD3 and CD28 stimulation when compared to cells transduced with empty vector prior to anergy induction (p<0.05). B. Additionally, these cells were analyzed for pERK generation by western blot after 30-minute stimulation with anti-CD3 and anti-CD28 coated beads.