Figure 1

ChX67779 activates the ISRE-luciferase reporter gene. (a) HEK-293 cells expressing luciferase under control of five interferon-stimulated response elements (ISRE) were incubated, at 4 × 10⁴ cells/well in a 96-well plate, with DMSO alone or increasing concentrations of ChX67779 ranging from 3 to 100 μM. After 24 hours of incubation, luciferase induction was determined. (b) HEK-293 cells were incubated with increasing concentrations of ChX67779 as described above. After 0, 24 and 48 hours of culture, the number of metabolically active cells was determined by ATP quantification using the CellTiter-GLO reagent. Results are expressed as a percentage relative to the initial number of living cells at t = 0 hour. (c) ISRE-luciferase reporter cells were left untreated (DMSO alone) or incubated with ChX67779 at 25 or 50 μM. Luciferase induction was determined after 4, 8, 16 and 24 hours of culture. (d) HEK-293T cells were transfected with pNF-κB-Luc or pISRE-Luc using 50 ng plasmid for 4 × 10⁴ cells/well in a 96-well plate. Cells were treated with ChX67779 at 50 μM or TNF-α at 10 ng/ml. After 24 hours of incubation, luciferase induction was determined. Experiments were performed in triplicate, and data represent means ± SD. *P < 0.05 and **P < 0.01 as calculated by one-way ANOVA with Tukey’s post hoc test (a and b), two-way ANOVA with Bonferroni’s post hoc test (c), or Student’s t test (d).