Figure 2.

Pre-TCR signaling is intact in Lck-FADD mice. (a) Flow cytometry was performed to analyze the percentage of cells in DN3a (lin⁻CD25⁺c-kit⁻CD27^medFSC^med) and DN3b populations (lin⁻CD25⁺c-kit⁻CD27^hiFSC^hi). Shown is a representative FACS plot to show gating strategy, with average DN3b percentage calculated from four Tg mice and three littermate controls also shown. Error bars reflect S.E.M. (b) Flow cytometry was performed to analyze expression of intracellular TCRβ (ic TCRβ) of DN2, DN3 and DN4 cells from Lck-FADD (up) or CD4-FADD (low) mice. Shown are representative data from four independent experiments. ***P<0.001. (c) FACS was performed to analyze expression of CD69 expression on the cell surface of DN3 and DN4 cells. Shown are representative data from three independent experiments. Error bars reflect S.E.M. ***P<0.001. (d) DN3, DN4 and DP thymocytes were gated as described in Figure 1. Cell surface CD5 expression and ic TCRβ expression were analyzed on all subsets. Shown are representative data from three independent experiments. ***P<0.001