Fig. 3.

Hsp90 inhibitors simultaneously block SMC3-induced NF-κB and Akt activation. a H23 and HepG2 cells were pretreated with 17AAG (50 nM) for 10 h, followed by SMC3 treatment (25 nM for H23 and 50 nM for HepG2 cells) for 24 h, IKKβ, RIP1, Bcl-xL and MnSOD proteins were detected by Western blot. β-tubulin was detected as an input control. b H23 and HepG2 cells were co-transfected with p5×κB-Luc and pRSV-LacZ. Twenty-four hours after transfection, the cells were pretreated with 17AAG (50 nM), CCT018159 (10 μM) or rifabutin (10 μM) for 10 h followed by SMC3 treatment for 24 h or left untreated. Luciferase activity was detected and normalized to β-galactosidase activity (n = 3). c H23 and HepG2 cells were pretreated with 17AAG (50 nM) for 10 h, followed by SMC3 treatment (25 nM for H23 and 50 nM for HepG2 cells) for 4 h. Phosphor-Akt (Ser⁴⁷³) and Akt were detected by Western blot. β-tubulin was detected as an input control