Figure 3

In vitro and in vivo effect of AS602868 combined with CPT-11/SN-38 on the NF-κB pathway. (A) IκB-α phosphorylation levels were studied by western blotting either on lysates of HT-29 cells that were stimulated 30 min with indicated concentrations of AS602868 and SN-38 or in tumours from mice treated as indicated. HSP60 was used as loading control. (B–C) NF-κB activation was visualized by EMSA. HT-29 cells were treated with indicated concentrations of AS602868, 30 min before stimulation with SN-38 (3 and 10 nM) for 1 h or with PMA (10 ng ml⁻¹) for 1 h as positive control. These results correspond to one representative experiment from 3. In supershift experiments, nuclear protein extracts of tumours from CPT-11 and AS602868±CPT-11-treated mice were incubated with anti-p50 and anti-p65 antibodies or rabbit IgG as negative control.