Protein and total RNA were extracted using Paris kit (Ambion-Life Technologies Ltd, Paisley, UK) according to the manufacturer’s instructions. Thirty μg of total protein from each lysate were boiled at 95 ºC for 5 min, separated by SDS/PAGE under reduced conditions (5% 2-mercaptoethanol) and transferred onto a nitrocellulose membrane. The membranes were subsequently blocked in 5% defatted milk-PBS for 1 h and incubated overnight at 4ºC with a primary antibody anti TRAP1 (1:1000, Labvision) or anti β-actin (1:10000, Sigma, Dorset, UK). Blots were then incubated with a horseradish peroxidase-linked secondary antibody (1:5000; Amersham Pharmacia Biotech, Little Chalfont, UK) and developed by chemoluminiscence with Lumilight plus kit (Roche diagnostics, Burgess Hill, UK). Apoptosis detection by western blotting was performed as described before (27 (link)).