AG14361 inhibits NF-κB activity and downregulates the levels of expression of NF-κB downstream genes. (A) Luciferase reporter assay for NF-κB transcription activity after treatment with IC50 of AG14361 for 24 hr in 92 J cell lines. (B) NF-κB transcription activity assay after treatment with IC50 of AG14361 and/or lestaurtinib for 24 hr in a HeLa cell line, which carries a stably integrated luciferase reporter. (C) The real-time PCR of five NF-κB downstream genes in 92 J cells. (D, E) Expression of IL6 in 92 J cells in the presence of AG14361 and various concentrations of lestaurtinib for 24 hr revealed by real-time PCR. (F-H) Expression of IL6 and COX2 in xenograft tumors after treatment of AG14361 and/or lestaurtinib. Data shown the mean of three different readings and are marked by columns with ± standard deviation (SD) error bars. IC50, 50% inhibition dose; NF-κB, nuclear factor κB.