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

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Janardhan et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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CARTg Th1 T cells were transduced with active Ras and preincubated for 30 minutes with varying amounts of the MEK-1 inhibitor PD98059. Cells were then either stimulated with ionomycin (50 mg/mL) for 30 minutes and analyzed for ERK phosphorylation by Western blot, or stimulated with ionomycin overnight in triplicate and supernatants were collected and analyzed for IL-2 production by ELISA. Star (*) indicates no detectable IL-2 produced by empty vector transduced cells stimulated with ionomycin. Error bars where shown indicate standard deviations of the mean between triplicate samples from the same experiment.