Relative activities conferred by AE1 and AE2.
A, schematic of the constructs assayed. B, Jurkat T cells in which CARD11 was stably knocked down were transfected with CSK-LacZ and Igκ2-IFN-LUC in the presence of expression vectors for the indicated Myc-tagged CARD11 variants and stimulated with anti-CD3/anti-CD28 cross-linking for 5 h as indicated. A two-tailed unpaired Student's t test with unequal variance resulted in p values <0.0085 for the values obtained under unstimulated conditions with the following constructs as compared with that observed with CARD11 ΔID: Δ441–546 Δ587–671, Δ441–654, Δ441–546, and Δ587–654. C, HEK293T cells were transfected with the same amounts of each expression vector used in B, and lysates were probed by Western blot (WB) using anti-Myc primary antibody to indicate the relative expression level of each variant. β-Galactosidase activity, driven by CSK-LacZ, was used to calculate equivalent amounts of lysate for Western analysis.