Fig. 7

Activation of the NFĸB pathway by serum in glioblastoma stem cells is associated with ROS stress. a–g Induction of expression of genes which are downstream of NFĸB by serum exposure. GSC11 cells were cultured in serum-free medium or serum-containing medium for 1, 3, and 7 days. The relative mRNA expression levels of the target genes were analyzed by microarray assays in triplicate for each time point. h Antioxidant NAC suppressed serum-induced NFĸB pathway activation in GSC23 cells. Protein samples were obtained from GSC23 cells cultured in serum-free or serum-containing medium in the presence or absence of 20 mM NAC for 3 days. The expression of phospho-IKBα, phosphor-p65, total p65, and β-actin was assayed by Western blot. i IKK inhibitor BMS-345541 and antioxidant NAC suppressed serum-induced NFĸB activation in GSC11 cells. GSC11 cells were cultured in serum-free or serum-containing medium with the indicated concentrations of BMS345541 or 20 mM of NAC for 3 days. The expression of phospho-IKBα, phosphor-p65, CD133, ANXA1, and β-actin was measured by Western blot analysis. CCND1 cyclin D1, CD44-4 CD44 transcript variant 4, CD44-5 CD44 transcript variant 5, FBS fetal bovine serum, GSC glioma stem cell, IL-8 interleukin 8, IL-11 interleukin 11, NAC N-acetyl-cysteine, NFĸB nuclear factor-kappa-B, PLAUR plasminogen activator urokinase receptor, ROS reactive oxygen species, TFP1-2 tissue factor pathway inhibitor 2