Fig. 5.

TMZ and PX-866 cooperate to diminish survival. a T98G (EGFR WT) and U373vIII (EGFRvIII) were treated with TMZ (500 μM) and increasing doses of PX-866 for 48 h (co-treatment). Both floating and remaining adherent cells were collected for analysis and western blotting for PARP and LC3 was used to assess apoptosis and autophagy, respectively. The triangle representing the dose escalation for PX-866 indicates: 0.125, 0.25, 0.5, and 1 μM. b T98G and U373vIII cells were treated with TMZ (500 μM) for 24 h, followed by the addition of PX-866 for another 24 h (sequential treatment). The triangle representing the dose escalation for PX-866 indicates: 0.125, 0.25, 0.5, and 1 μM. T98G cells (c) and U373vIII cells (d) were treated with the indicated treatments for 24 h. Cells were trypsinized 24 h after drug treatment and re-plated in 35 mm dishes in triplicate and allowed to form colonies. After about 9–10 days, colonies were fixed (6% glutaraldehyde) and stained (0.5% crystal violet). The number of colonies were counted and presented as a bar graph. Values are the mean ± the standard deviation of three separate measurements, *, P < 0.05 and, **, P < 0.01. Data represents the mean and S.D. from three independent experiments with each experiment conducted in triplicates