Figure 3.

Smac mimetic activates the NF-κB pathway. (a) GBM10 cells were treated for indicated times with 1 μM BV6 or DMSO; stimulation with 10 ng/ml TNFα for 1 hour was used as positive control. Nuclear extracts were analyzed for NF-κB DNA-binding activity by EMSA. A representative experiment of two experiments is shown. (b) GBM CSLCs were treated for indicated times with BV6 (GBM9: 0.6 μM, GBM10: 1 μM BV6) or DMSO. Expression levels of p100 and p52 were analyzed by western blotting. Expression level of GAPDH served as loading control. (c) GBM CSLCs were treated for 7 days with BV6 (GBM9: 0.6 μM, GBM10: 1 μM BV6) or DMSO; stimulation with 10 ng/ml TNFα for 1 hour was used as positive control. Expression levels of p100, p52, p65 and p50 were analyzed in cytoplasmic (C) and nuclear (N) fractions by western blotting. α-tubulin served as purity and loading control for cytoplasmic fractions and lamin A/C for nuclear fractions