Figure 5.

MIP-1α co-induction by palmitate and TNF-α implicates signaling via c-Jun/NF-κB. To test whether the MIP-1α co-induction by palmitate and TNF-α involved c-Jun and NF-κB signaling, THP-1 monocytic cells were treated with palmitate (200 μM for 1 h) and TNF-α (10 ng/mL for 10 min) while control wells were treated with vehicle only. Cells were lysed in RIPA buffer and 20 μg protein from each sample was loaded for analysis by SDS-PAGE. The representative data (mean ± SEM) from three independent determinations with similar results show the downmodulated expression of (A) MIP-1α transcripts (P = 0.01) and (B) MIP-1α secretory protein (P = 0.002) in resveratrol-treated cells as compared to controls treated with the vehicle only before co-stimulation with palmitate and TNF-α. (A) Western blots show the expression of hyperphosphorylated c-Jun (p-cJun) and NF-κB (p-NF-κB) proteins against respective total proteins in THP-1 monocytic cells co-stimulated with palmitate and TNF-α as compared to their expression in cells treated with either palmitate or TNF-α. Normalization of phosphorylated against total proteins shows significantly increased expression of (B) phosphorylated c-Jun and (C) phosphorylated NF-κB in cells co-stimulated with palmitate and TNF-α, compared to individual stimulations with palmitate or TNF-α. * P < 0.05 and ** P < 0.01.