Figure 1: JKAP deficiency enhances T-cell activation and TCR signalling.

(a) Flow cytometry analyses of T-cell activation markers CD69 (0–1 μg ml⁻¹, 12 h; left panel) and CD25 (0.5 μg ml⁻¹, 24–72 h; right panel) on murine splenic T cells on anti-CD3 stimulation. (b) Carboxyfluorescein diacetate succinimidyl diester dilution assays of T-cell proliferation at 48 h (left panel) and 72 h (right panel) on anti-CD3 (0.5 μg ml⁻¹) stimulation. (c) ELISAs of IL-2, IFN-γ and IL-4 levels in culture supernatants from splenic T cells treated with anti-CD3 (0–1 μg ml⁻¹) stimulation or anti-CD3 (0.5 μg ml⁻¹) plus anti-CD28 (0.2 μg ml⁻¹) costimulation for 24 h. (d) Immunoblotting of various TCR signaling molecules in purified splenic T cells on anti-CD3 (5 μg ml⁻¹) stimulation. (e) Immunoblotting of the phosphorylation of IKK and ERK in splenic T cells on phorbol myristate acetate plus ionomycin (P+I) treatment. Relative fold changes were normalized to total protein levels and are shown at the bottom of the panel (d,e). Quantification of p-Lck (Y394) and p-ZAP-70 (Y493) levels is shown at the bottom of the panel (d). Data are presented as means±s.d. Two-tailed Student’s t-test, *P<0.05. Data are representative of at least three independent experiments. Prol. Ind., proliferation index; WT, wild-type; KO, JKAP-knockout.