Chemicals

Acrylamide, bovine serum albumin (BSA), bromophenol blue, DMBA, ethidium bromide, 2-mercaptoethanol, sodium dodecyl sulphate (SDS), N,N,N’,N’-tetramethylene diamine (TEMED) and trizol were purchased from Sigma Chemical Company, St. Louis, MO, USA. Power SYBR^® Green PCR master mix was obtained from Applied Biosystems, California, USA. Antibodies for cyclin D1, CDK4, p21, p53, GSK-3β, p-GSK-3β^Tyr216, Akt, PI3K, cleaved caspase-3, cleaved caspase-9, cytochrome c, Opa-1 and Gapdh were procured from Santa Cruz Biotechnology, USA. Antibodies for Bcl-2, Bax, p-Akt^Ser473, β-catenin, p-β-catenin^{Ser33,Ser37,Thr41}, p-β-catenin^Ser552, CREB, p-GSK-3β^Ser9, ERK1, p-ERK1^{Thr202/Tyr204}, p-cyclin D1^Thr286, cleaved PARP and histone as well as ELISA kits were from Cell Signaling Technology, USA. Oligonucleotide primers and primers for mature microRNA were procured from Sigma Genosys, San Ramon, USA (The primer details are provided in Supplementary Table S4). Nimbolide was obtained from M/s Asthagiri Herbal Research Foundation, Chennai, India. Wortmannin was obtained from Selleckchem, USA. All other reagents used were of analytical grade.