Ectopic overexpression of E127G CARD11 in normal T cells reduces magnitude of AgR-induced response. (A) Representative expression of endogenous or GFP-fused CARD11 in normal T cells transfected with empty vector (EV), WT, or E127G CARD11-GFP. (B) Representative flow cytometric analysis of CD69 expression on transfected cells stimulated as in Fig. 6 C. MFI values are listed for each histogram. (C) IL-2 secretion measured by ELISA in transfected cell supernatants 3 d after stimulation as described in Fig. 6 C. Data show mean ± standard deviation of triplicate wells. (D) Fold induction of CD69 expression and IL-2 secretion was calculated by normalizing CD69 MFI or IL-2 concentration in stimulated cells to unstimulated cells for each transfection. (E) Representative proliferation, as measured by dilution of CellTrace Violet dye, of unstimulated and bead-stimulated transfected T cells. The marker denotes the percentage of T cells that divided five to seven times. (B–E) Data are representative of two independent experiments.