Fig. 4.

Early T-cell activation requires Akt phosphorylation. a Immunoblot for pAkt Thr308 and Ser473 and β-actin in NV, EM and CM T-cells following 0, 15, 30 and 180 min of activation with anti-CD3/CD28. Densitometry of pAkt Thr308 (b) and pAkt Ser473 (c) for NV, EM and CM T-cells normalised to β-actin. ECAR (d) and OCR (e) in NV CD4+ T-cell upon treatment with either 10 μM Akt1/2 kinase inhibitor or vehicle control (DMSO) and anti-CD3/CD28 at the times indicated. Data are representative (a–c) of 3–5 experiments with one representative immunoblot sample of 3–5 is shown, (d, e) three experiments. Statistical analysis was performed using a nonmatching two-way ANOVA with Sidak’s multiple comparison test (b, c). Data are expressed as mean ± SEM; **p ≤ 0.01