Effect of GS12021 treatment on LPS-stimulated cell signaling in RAW 264.7 cells.

<p>Immunoblot analyses of phosphorylated IKK, IκBα and phosphorylated- and total-p65. The experiments were repeated at least three times, and the representative images and the densitometry results for phosphorylated p65 are shown in (A). Phosphorylations of JNK, ERK, p38 MAPK, Akt, c-Jun and S6 protein (B). Cells were pretreated with vehicle or GS12021 (20 μM for 1 h) and stimulated with LPS for the time periods indicated. (C) NF-κB reporter assay (N = 4). (D) AP-1 reporter assay (N = 4). RAW 264.7 cells were transfected with NF-κB or AP-1 luciferase reporter plasmid and, after 24 h, cells were incubated with GS12021 (20 μM or 40 μM) for a further 24 h. **<i>P</i> < 0.01 <i>versus</i> CON; ^##<i>P</i> < 0.05 <i>versus</i> LPS alone.