Figure 4.

Effects of juglone on IL-34-induced AP-1 activity and cell transformation in JB6 Cl41 cells. Cells were seeded and co-transfected with the luciferase reporters, c-Fos-luc (A), c-Jun-luc (B), AP-1-luc (C) along with pRL-TK vector. At 24 h after transfection, the cells were serum starved for 24 h, and then treated with the indicated doses of IL-34 for 24 h before a luciferase assay was performed. (D) Cells were seeded and co-transfected with the luciferase reporter AP-1-luc and pRL-TK vector. After 24 h, the cells were serum starved for 24 h, pretreated with the indicated doses of juglone and then exposed to 10 ng/mL IL-34 for 24 h before a luciferase assay was performed. (E) Cells were treated with 10 ng/mL IL-34 in the presence or absence of juglone in a soft agar matrix and incubated at 37 °C in a 5% CO₂ atmosphere for 14 days. Representative colonies from three separate experiments were photographed (left), followed by calculation of the average colony numbers and sizes (diameter > 200 μm, right). Data in (A–E) represents the mean ± S.D. of triplicate measurements from three independent experiments. Statistical analyses were conducted using one-way ANOVA (*p < 0.05, ** p < 0.01, *** p < 0.001, compared to control group or only IL-34-treated group).