Figure 3. EET inhibits the NF-κB pathway.

NP cells were cultured in serum-free media for 8 hours. Then, the cells were treated with EET (2μM) or vehicle for 2 hours. Subsequently, TNF-α (50 ng/ml) was added, and cells were incubated for 10, 30, or 60 minutes. Cells were then collected and total protein extracted. The expressions of p-p65, p-p38, p-ERK, p-JNK, and p-AKT were detected by immunoblotting. Antibodies to GAPDH and total p65, p38, ERK, JNK, and AKT were used as loading controls. The immunoblots were quantified using ImageJ software. Experiments were performed in triplicate.