NF-B/AP-1-Targeted Inhibition of Macrophage-Mediated Inflammatory Responses by Depigmenting Compound AP736 Derived from Natural 1,3-Diphenylpropane Skeleton

Figure 4

Effects of AP736 on the transactivation activity of transcription factors and their nuclear translocation. ((a) and (b)) HEK293 cells cotransfected with NF-κB-Luc and AP-1-Luc constructs, as well as a β-gal construct as a transfection control, were treated with AP736 upon cotransfection with MyD88 or TRIF (1 μg/mL each) for 8 h. The resultant luciferase activities were determined using luminometry as described in Section 2. (c) RAW264.7 cells (5 × 10⁶ cells/mL) were incubated with LPS (1 μg/mL) in either the presence or absence of AP736 for the indicated times. After preparing nuclear fractions, the translocated levels of total or phosphorylated transcription factors (p65, p50, c-Fos, c-Jun, and lamin A/C) were identified using immunoblotting. and compared with the control group.

(a)

(b)

(c)