Fig. 5

Alterations of signaling pathways by synergistic combination treatments in glioma tumor-initiating cells. a Cells were treated with vehicle as control (C), erlotinib (E), sorafenib (S), U0126 (U), and combinations for indicated duration, and 10 μg of extracted proteins in each sample were subjected to immunoblotting using indicated antibodies to detect total or phosphorylated (p-) proteins. Concentrations of drugs were as follows: erlotinib; 0.6 μM for GSC11 and 6 μM for GSC20, sorafenib; 4 μM for GSC11, U0126; 10 μM for GSC11 and 20 μM for GSC20. β-actin was examined as loading control. b GSC11 or GSC20 cells were treated with indicated reagents for 24 h and 10 g of extracted nuclear or cytoplasmic proteins in each sample were subjected to immunoblotting using anti-PKM2 and anti-β-catenin antibodies. Lamin B and vinculin were examined as loading controls of nuclear proteins and cytoplasmic proteins, respectively. Relative values of blots of PKM2 and β-catenin in treatment groups to control were calculated using ImageJ