Fig. 6

Ibrutinib inhibits EGFR-induced NF-κB activation in glioma cells. (a) The cells were treated with indicated concentrations of ibrutinib for 3 h. The whole cell extracts were analyzed by Western blot analysis using IκB-α antibody. (b) Ibrutinib suppressed EGF-induced degradation of IκB-α. U87 and U251 cells were pretreated with or without ibrutinib (30 μM) for 2 h. The cells were then stimulated with EGF (100 ng/mL) for the indicated times. The expression of IκB-α was analyzed by Western blot analysis. (c) Ibrutinib inhibited nuclear translocation of p65 in glioma cells. The cells were treated with indicated concentrations of ibrutinib for 3 h. The nuclear extracts (NE) and cytoplasmic extracts (CE) were analyzed by Western blotting. (d) Ibrutinib inhibited EGF-induced nuclear translocation of p65 in glioma cells. U87 and U251 cells were pretreated with or without ibrutinib (30 μM) for 2 h. The cells were then stimulated with EGF (100 ng/mL) for the indicated times. The cytoplasmic and nuclear protein extracts were analyzed by Western blotting. (e) Ibrutinib inhibited degradation of IκB-α and nuclear translocation of p65 in U87 EGFRvIII cells. (f) BTK knockdown cells were stimulated with EGF (100 ng/mL) for indicated time and then cell lysates were subjected to immunoblotting analysis using indicated antibodies