Phospho p70s6 kinase thr

24 h after plating, 50% confluent cells were treated. The drug concentrations applied are listed in Table 3 (single treatment) respectively Table 4 (combination treatment). After 6 h, 24 h or 72 h for single or combination treatment, respectively at day 8 of the multimodal RIST and TIST treatment, the cells were harvested. The whole cell lysates were separated by gel electrophoresis and the proteins were transferred to nitrocellulose membranes (Amersham^TM Protran^TM 0,45 µm, GE Healthcare Life Science, Cat. No. 10600002). After the transfer, the membranes were incubated with primary antibodies (1:1000, respectively 1:2000 for LC3B and GAPDH antibody). The primary antibodies AKT (pan, #4691), phospho-AKT (Ser473, #4060), phospho-AKT (Thr308, #13038), 4E-BP1 (#9644), phospho-4E-BP1 (Ser65, #9456), MycN (#9405), p70S6 kinase (#9202), phospho-p70S6 kinase (Thr389, #9234), Src kinase (#2108), phospho-Src family (Tyr416, #2101), caspase-3 (#9665) and p21 Waf1/Cip (DCS60, #2946) were purchased from Cell Signaling. The LC3B antibody (#NB100-2220SS) was purchased from NovusBio, the PARP-1 antibody (#1072-1) was purchased from Epitomics and the GAPDH antibody (#Sc-47724) was purchased from Santa Cruz. ImageJ was used for quantification of the protein expression.