Fig. 6.

Thiocolchicoside represses NF-κB–dependent reporter gene expression induced by TNF and by overexpression of various signaling intermediates. A, thiocolchicoside inhibits the NF-κB–dependent reporter gene expression induced by TNF. A293 cells were transiently transfected with a NF-κB–containing plasmid for 24 h. After transfection, the cells were incubated with the indicated concentrations of thiocolchicoside for 24 h and then treated with 1 nmol/L TNF for an additional 24 h. The supernatants of the culture media were assayed for SEAP activity. Data are presented as mean (±SD). B, thiocolchicoside inhibits the NF-κB–dependent reporter gene expression induced by TNF, TNFR1, TRADD, TRAF2, NIK, IKK, and p65. Cells were transiently transfected with a NF-κB–containing plasmid alone or with the indicated plasmids. After transfection, cells were incubated with 100 μmol/L thiocolchicoside for 24 h and then incubated with the relevant plasmid for an additional 24 h. TNF-treated cells were incubated with 100 μmol/L thiocolchicoside for 24 h and then treated with 1 nmol/L TNF for an additional 24 h. The supernatants of the culture media were assayed for SEAP activity. Data are presented as mean (±SD). C, thiocolchicoside inhibits the COX-2 promoter activity induced by TNF. Cells were transiently transfected with a COX-2 promoter linked to the luciferase reporter gene plasmid for 24 h and treated with the indicated concentrations of thiocolchicoside for 24 h. Cells were then treated with 1 nmol/L TNF for an additional 24 h, lysed, and subjected to a luciferase assay. Data are presented as mean (±SD), and * indicates P < 0.05 when compared with their respective control.