Resveratrol inhibits NF-κB signaling in DC exposed to TNF-α.

<p>DC were stimulated with TNF-α, in presence or absence of resveratrol, for the indicated times. <b>A.</b> Cells were stained for NF-κB (p65 subunit, green staining) and nuclei were counterstained with DRAQ5 (red staining), before acquisition by ImageStream^X. Representative plot of the similarity coefficient between nucleus and NF-κB stainings of unstimulated DC (grey), DC stimulated with TNF-α alone (red), or in presence of resveratrol (Rsv, green). The yellow line corresponds to the gate for which nuclear translocation of NF-κB was considered. Three representative images of cells found in the indicated areas of the graph are shown. For the control condition, cells with a 0 similarity coefficient (no p65 translocation, grey line), and for the TNF-α -stimulated condition, cells with a similarity coefficient of 1 (NF-κB nuclear translocation, red line) are shown. BF: brightfield. <b>B.</b> Quantification of the percentage of cells with nuclear NF-κB along time (within the translocated gate) for different donors. Each symbol represents a different individual (the mean value is represented as a dash), and TNF-α stimulation in absence or presence of resveratrol is represented by closed or open symbols, respectively. Comparisons were performed using Friedman test, followed by Dunns multiple comparisons test (* p<0.05). <b>C.</b> Phosphorylation of p65 in response to TNF-α, in presence or absence of resveratrol or BAY 11-7082 (BAY) and <b>D.</b> MAPK signaling in response to TNF-α and/or resveratrol was evaluated by Western blot. DC were stimulated with TNF-α, in presence or absence of resveratrol, for indicated times. Cells were lysed and samples resolved in reducing gels, which were blotted onto nitrocellulose membranes, and probed with the indicated antibodies. The images are representative of three independent experiments. p: phosphorylated.