Fig. 2.

TL +/− Btz downregulates cIAP1/2 but does not inactivate the canonical NF-κB pathway. a U266 and PS-R cells were treated with Btz +/− TL for 24 h, after which cIAP1 and cIAP2 were monitored by immunoblotting analysis. α-Tubulin was assayed to ensure equivalent loading and transfer. b U266 cells were exposed to the indicated concentrations of Btz +/− TL for 16 h, after which nuclear protein was extracted from the cells. Immunoblotting analysis was then performed to monitor levels of p65. p84 was assayed to ensure equivalent loading and transfer. c U266 cells were incubated with 500 nM TL32711 +/− 3 nM Btz for 4 h, 8 h, and 16 h, after which p-p65 (S536) was monitored by immunoblotting analysis. β-actin was assayed to ensure equivalent loading and transfer. d Following treatment as in b, nuclear proteins were isolated using a Nuclear Extract Kit. DNA binding of NF-κB (p65 subunit) was determined using TransAM for NF-κB activity. ***P < 0.001; NS not significant