The regulation of NF-kB by heptadecane via NIK/IKK and MAPKs induced by oxidative stress.

Abstract

<p>YPEN-1 cells were grown until 80% confluent in 100 mm dishes in DMEM medium. Cells were pre-treated (1 hr) with heptadecane (1, 10, or 20 µM) or NAC (2 mM), then stimulated with 10 µM t-BHP. (A) After stimulation with t-BHP (1 hr for NF-kB) in the absence (-) or presence (+) of heptadecane (1, 10, or 20 µM), cells lysed and total nuclear and cytosolic proteins were extracted. Western blot was performed for p50, p65 and IκB. (B) After stimulation with t-BHP (10 min for phosphorylated NIK and 20 min for phosphorylated IKK) in the absence (-) or presence (+) of heptadecane (1, 10 or 20 µM), cells were lysed and p-NIK and p-IKKα/β levels were determined. (C) After stimulation with t-BHP (10 min for phosphorylated MEK1/2 and 20 min for phosphorylated MAPKs) in the absence (-) or presence (+) of heptadecane (1, 10, or 20 µM), cells were lysed and p-MEK1/2, p-ERK1/2, p-p38, and p-JNK levels were determined. One representative experimental blot of each protein is shown from three experiments that yielded similar results.</p