Fig. 4.

IKK-target genes are up-regulated in primary tumors and inhibition of IKK reduces tumor volume in vivo. (A) ChIP with α-SMRT antibody and PCR detection of the hes1 and hes5 gene and mRNA expression by semiquantitative RT-PCR from HCT-116, HT29M6, and SW480 treated with DMSO or BAY11-7082 for 48 h. (B) Semiquantitative RT-PCR showing the expression levels of hes1 and hrt1/herp2 in SW480 cells transfected with control vector or DN-IKKα. (C) Representative culture of SW480 cells after 3-day treatment with DMSO or BAY11-7082. (Left) Graphs represent daily cell counts from cultures treated with the indicated concentrations of BAY11-7082. (Center) Photographs show a representative image of the cultures at day 3 of treatment. Images were obtained in an Olympus IX-10 at ×100. (Right) Flow cytometry analysis of AnnexinV staining. (D) CRC-X8 and CRC-X34 tumors were implanted s.c. in nude mice (n = 40). Ten days after transplantation, the animals were treated with BAY11-7082 or DMSO as a control. Tumors were measured weekly, and the results are presented as mean ± SEM. Student's t test for independent analysis was applied to evaluate differences in tumor volume [A(B²)π/6] between treated and control mice.