250

1. > 2 antagonism 1 additivity synergy 0 BEZ235 (nM) 5 7.5 7.5 7.5 7.5 7.5 7.5 7.5 7.5 5 5 5 5 5 5 5 TXT (nM) 0.1 0.5 0.5 0.5 0.5 0.1 0.5 0.5 0.5 0.1 0.1 0.5 0.1 0.1 0.1 0.1 Carboplatin (mM) 250 100 250 100 250 100 100 250 100 100 100 250 250 250 250 100 A HS578T HS578T B 120 100 80 MTT Metabolization (% Control) 60 40 20 0 TXT B +T B +C Control Control BEZ235 BEZ235 1st T / 2nd B 1st B / 2nd T 1st C / 2nd B 1st B / 2nd C Carboplatin Supplementary Figure 6. Effect of the combination of BEZ235 and standard of care drugs on HS578T. A, Cells were treated with the indicated concentrations of the drugs, either in double or triple combinations, and their MTT values were measured. Combination indexes for the different drug combinations were obtained using the CalcuSyn program and plotted. B, Effect of sequential versus concomitant administration of the drug combinations on MTT metabolization rates. Cells were plated, and 24 hours later treated with the individual drugs (BEZ235, 7.5 nM; Carboplatin, 100 mM; or Taxotere, 0.5 nM) or the drug combinations (B + C, or B + T) for 72 hours, in the case of concomitant administration. For the sequential treatments, the first drug was added at the beginning of the experiment, and the second drug was added 15 hours later. MTT metabolization was analyzed in all cases at 72 hours. The results represent the mean ± SD of quadruplicates of an experiment which was repeated twice. Supplementary Figure 6