Hrp anti mouse

Etoposide (cat# E1383), Nicotinamide (cat# 479865-U) and Sirtinol (cat# s7942) were from Sigma-Aldrich Company Ltd (Dorset, UK). Cisplatin (cat# 440-040) and metothrexate (cat# 440-045) were from Enzo Life Sciences UK Ltd (Exeter, UK). EX527 (cat# 2780) was from Tocris bioscience (R&D systems, UK). Tissue cell culture media were supplied by Gibco Life Technologies and foetal calf serum by Harlan Seralab (UK). Cyclophilin A (cat# Ab3563), actin (cat# Ab8226), SIRT1 antibody (cat# Ab13749), and HRP-anti-mouse (cat# Ab6808) antibodies were from Abcam (UK). p53 BC-12 antibody (cat# SC126) was from Santa-Cruz Biotechnology (Texas, USA). c-Myc antibody (cat# 9402), Bcl-2 (cat# 2870) and HRP-anti-rabbit (cat# 7074) antibody were from Cell Signalling Technology (MA, USA). miR-34a mimic (cat# C-300551-07), control (cat# CP001000-02-05) were purchased from Dharmacon (now GE Healthcare, UK). siRNAs targeted for p53 (cat# 1299001) and scrambled siRNA were from Invitrogen.

Third-instar larval brains were lysed in RIPA lysis buffer [50 mM Tris-HCl pH 8.0, 150 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS, 1% IGEPAL CA-630 and complete protease inhibitor cocktail (Roche)]. About 30 brains were lysed in each sample. Samples were subjected to SDS‐PAGE and transferred to a polyvinylidene fluoride membrane. Mouse anti-His3 (1:1000; Active Motif) and mouse anti-β-Tubulin (1:1000; DSHB) were used as primary antibodies, and HRP, anti‐mouse (1:5000; Abcam) were used as secondary antibodies.