4-Isopropyl-2,6-bis(1-phenylethyl)aniline 1, an Analogue of KTH-13 Isolated from Cordyceps bassiana, Inhibits the NF-κB-Mediated Inflammatory Response

Figure 2

Effects of KTH-13-AD1 on transcriptional activation of LPS-mediated inflammatory responses. (a, b) mRNA levels of proinflammatory genes (iNOS and TNF-α) in LPS-treated RAW264.7 cells pretreated with KTH-13-AD1 (150 μM), as analyzed by RT-PCR (a) and real-time PCR (b). (c, d) Transcriptional activation of inflammatory responses was examined using a luciferase reporter gene assay with HEK293 cells transfected with NF-κB-Luc or AP-1-Luc, as well as with β-gal (as a transfection control) under either treatment with PMA or cotransfection with FLAG-MyD88 or CFP-TRIF (1 μg/mL each). Luciferase activity was measured using a luminometer. (e) The nuclear translocation levels of AP-1 (c-Jun and c-Fos) and the NF-κB (p65 and p50) family in LPS-treated RAW264.7 cells (5 × 10⁶ cells/mL) pretreated with KTH-13-AD1, as examined using nuclear fractionation and immunoblotting analysis. compared to the vehicle control.

(a)

(b)

(c)

(d)

(e)