Examples of Akt Thr308 and Ser473 phosphorylation, together with the phosphorylation of downstream substrates. Triplicate lysates of normal (N1–3) and patient-matched tumour (T1–3) tissues were separated on SDS–PAGE gels. Phosphorylation of Akt on Thr308 and Ser473, and of three downstream substrates, was determined by western blotting with phospho-specific antibodies as indicated. An anti-F1-ATPase antibody was used as a control for protein loading. (A) Examples (patients 11 and 24) of tumours in group Ia. (B) Examples of tumours from two patients (patients 17 and 27) in group Ib. (C) Examples of two patients (patients 4 and 7) in group II.