Anti gadd45α

Transfection reagents Lipofectamine 2000 and Lipofectamine 3000 were purchased from Invitrogen, RNA synthesis inhibitor actinomycin D, protease inhibitor MG132, lysosomal inhibitors bafilomycin and hydroxychloroquine, CDK inhibitor RO-3306, and thymidine were purchased from Sigma-Aldrich. λPPase was purchased from New England BioLabs. Anti-CELF6 (ab173282) and anti-GFP (ab1218) were obtained from Abcam, anti-p21^Waf1/Cip1 (#2947), anti-p27^Kip1(#3686), anti-Gadd45α (#4632), anti-Cyclin B1(#12231), anti-β-TrCP (#4394), anti-Wee1 (#13084), anti-cleaved PARP (#5625), anti-cleaved Casp3 (#9661), anti-cyclin E1 (#20808), and anti-Ubiquitin (#3936) antibodies were purchased from Cell Signaling Technology, anti-P53 (10442-1-AP), anti-GFP (66002-1-1 g), anti-His (66005-1-1 g), anti-β-tubulin (60008-1-1 g), and anti-GAPDH (10494-1-AP) were obtained from Proteintech, anti-LC3B (L7543) was obtained from Sigma-Aldrich.

The chemotherapy compounds mitoxantrone (MX) (Wyeth-Lederle, Finland), doxorubicin (DXR) (Nycomed, Roskilde, Denmark), cyclophosphamide (Orion Pharma, Espoo, Finland), topotecan (GlaxoSmithKline, Uxbridge, Middlesex, UK), etoposide (Pfizer), cisplatin (Bristol-Myers Squibb, Princeton, NJ, USA), docetaxel (Aventis), and carboplatin (Bristol-Myers Squibb, Princeton, NJ, USA) were stored and prepared as described [17] (link). Imatinib was a gift from Novartis Pharmaceuticals (Basel, Switzerland). The stock solution of imatinib at 200 mM was prepared by dissolving the compound in DMSO. The c-kit and PDGF-α and β receptor blocker AG1296 was purchased from Calbiochem (cat. no. 658551). PDGF-BB was purchased from Sigma-Aldrich. The following antibodies were used for Western blotting: mouse monoclonal DO-1 for p53 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-GADD45α (Cell Signaling, cat. no. 3518), rabbit polyclonal anti-phospho-PDGF β receptor (Cell Signaling, cat. no. 3161), monoclonal mouse anti-RAD51 (Invitrogen, Carlsbad, CA, USA; cat. No. 35–6500), monoclonal mouse anti-cyclin B1 (BD Biosciences, cat. no. 554178) and monoclonal anti-p73 (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The RNA was isolated using the RNeasy kit (Qiagen, Hilden, Germany).