Figure 10
Inhibition of pAKT augments HSP27 siRNA suppression of survival but does not alter sensitivity to temozolomide (TMZ) in primary glioma cell lines. A. Representative Western blots of HF373 and HF2303 cells grown in serum-free medium (SF DMEM) or treated with control siRNA (Csi) or HSP27 siRNA (HSP27si) for 72 hr. B. Representative Western blots of HF2303 cells treated with control siRNA (Csi) or HSP27 siRNA (HSP27si) for 72 hr. Equal numbers of siRNA-treated cells were plated overnight in growth serum then treated ± AKT inhibitor IV at the concentrations indicated for 2 days before lysing. Arrows indicate ≥ 2-fold increases or decreases due to siRNA treatment. # - Indicates a ≥ 2-fold increase in the ratio of LC-3II/LC-3I. Asterisks indicate ≥ 2-fold increases or decreases due to AKT inhibitor IV treatment. C. Means for surviving fraction ± SD for HF2303 cells treated with Control (Csi) or HSP27 (HSP27si) siRNAs ± AKT inhibitor IV plating 3000 cells/60-mm dish. * p = 0.0061, ** p = 0.0008. D. Means for surviving fraction ± SD for HF203 cells treated ± AKT inhibitor IV ± TMZ plating 3000 cells/60-mm dish. * p = 0.029. E. Means for colony forming efficiency ± SD for HF2303 cells treated with Csi or HSP27si siRNAs ± AKT inhibitor IV ± TMZ plating 3000 cells/60-mm dish. * p = 0.017, ** p = 0.0005, *** p = 0.0001, and **** 0.0012. Plating 6000 cells/60-mm dish gave similar results.