For individual produced tumor grafts consent for tumor use was obtained from patients under a project approved by the Vall dHebron Hospital Clinical Investigation Ethical Committee. Tumors were subcutaneously implanted in 6 week-old female HsdCpb: NMRI Foxn1nu mice. Animals were supplemented with 1uM estradiol within the drinking-water. After tumor graft growth, Imatinib Gleevec tumor tissue was re-implanted in to recipient mice, of randomized upon growth. FDG Dog Checking 0. 3 to 0. 4 mCi of fluorine 18 deoxyglucose were injected intravenously through the vein of the anesthetized mouse. After having a period of 1 hour the mouse was imaged over a NanoPET/CT scanner. The NanoPET/CT is a highresolution little dog multi-modality scanner consisting of 12 lutetium yttrium oxyorthosilicate sensor blocks. The blocks comprise a total of 39,780 crystals each with a aspect of 13 mm3. Images were obtained in three-dimensions. The rats remained supine and maintained their position throughout the procedure. First, a CT scan was performed and second, Cellular differentiation a whole-body 18F FDG PET exhaust scan was obtained since the same area as the CT scan. Counts per minute were obtained, transformed into mCi, and values were normalized for ROI volume and injected dose. So that you can correct for metabolic variability between examinations and to determine growth particular uptake improvements, FDG uptake rates were adjusted for cardiac FDG uptake. For studies involving repeat checking, the change in growth specific FDG uptake was determined in percent 100. Animals were housed in the Longwood SAIF satellite dog facility between tests. Immunohistochemistry For immunohistochemistry we used anti cleaved buy Tipifarnib caspase 3, anti Ki67. Other antibodies used are described within the immunoblotting area below. All immunohistochemistries were done as explained previously including antigen collection using a citrate buffer. Immunoblotting Cells were treated with mock, NVP BKM120, Olaparib, KU 55933 or even the mixture and lysed in cell lysis buffer depending on the manufacturers instructions. Immunoblots were performed using the Nupage System. A complete of 20 ug of protein were loaded, apart from PAR, Phospho ATM and Phospho DNA PK/PRKDC american blots, where 40 ug were loaded. Cancer tissue lysates were prepared equally with the exception of tissue homogenization by using an electrical homogenizer for 30 secs after addition of the lysis buffer. Primary antibodies used for western blotting were total AKT, Cleaved Caspace 3, total ERK, Phospho AKT Ser473, Phospho ERK Thr202/Tyr204, Phospho Histone H2AX Ser139, PTEN from Cell-signaling. Phospho ATM Ser1981, Phospho DNA PK/PRKDC Ser2056 from Epitomics, Inc. CD31, Actin, INPP4B from Abcam, pADPr from Santacruz Biotechnology, and Ki 67 was bought from Thermo Scientific. Rad51 antibody was something special from Dr. Ralph Scully.