Nvp bkm120

NVP-BEZ235 (BEZ235), NVP-BKM120 (Buparlisib, BKM120), GSK1120212 (Trametinib, JTP-74057), PD0325901, and AZD6244 were purchased from Chemie Tek (Indianapolis, IN). Temozolomide, doxorubicin and paclitaxel were purchased from Sigma. For in vivo studies, BEZ235 and BKM120 were resuspended in NMP (N-methyl-2-pyrrolidone):PEG300 1:9 v:v, and GSK1120212 and PD0325901 were resuspended in 0.5% hydroxypropyl-methylcellulose (Sigma):0.2% Tween 80 (Sigma) (Gilmartin et al., 2011 (link); Kinross et al., 2011 (link)). BKM120, GSK1120212 and PD0325901 were given once daily by oral gavage at 10 ml/kg. Mice were treated with BEZ235 on a 5 days on, 2 days off schedule (Liu et al., 2009 (link)). Vehicle- and drug-treated mice were closely monitored on a daily basis for clinical signs as described above.

Soluble human recombinant SuperKillerTRAIL (referred to as TRAIL in this study) was purchased from Enzo Biochemicals (Enzo Life Sciences, Farmingdale, NY, USA). NVP-BKM120 was purchased from Chemie Tek (Indianapolis, IN, USA). Caspase inhibitors (z-VAD-fmk, z-IETD-fmk, z-DEVD-fmk, and z-LEHD-fmk) were purchased from R&D Systems (Minneapolis, MN, USA). The following antibodies were used: Mcl-1 (#4572), cytochrome c (#4280), cleaved poly-ADP-ribose polymerase (PARP, #9546), cleaved caspase-3 (#9664), cleaved caspase-8 (#9496), cleaved caspase-9 (#9501), Smac/DIABLO (#2954), pAKT (#5106), AKT (#2920), pS6kinase (#5364), pBAD (#9105), BIM (#2819), Bcl-2 (#2872), Bcl-xL (#2764), surviving (#2808), XIAP (#2042), DcR1 (#4756) and β-actin (#4970) were from Cell Signaling Technology (Beverly, MA, USA). Noxa (sc-26917) and apoptosis-inducing factor (AIF; sc-5586) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). DR4 (#IMG-141A) and DR5 (#IMG-122A) were from Imgenex (San Diego, CA, USA).