YC-1 Impairs VEGF-Induced Nuclear Translocation of NFκB. A–F. Immunofluorescence Analysis.

<p>hRMVECs were incubated for 8 hours under various conditions and were subsequently fixed for immunocytochemistry. The staining intensity and the subcellular localization of NFκB/p65 were determined by immunofluorescence microscopy using anti-NFκB/p65 antibody. Photomicrographs, which exhibit intense nuclear and/or cytoplasm staining was considered a positive signal. Cells that were stimulated with VEGF alone (B), or incubated with either VEGF/SN50M (C) or VEGF/DMSO (D); exhibited high levels of NFκB/p65 immunoreactivity, which was preferentially localized in the nuclei of the cells. In addition, there was a positive intense staining signal of NFκB/p65 deposited over the cytoplasms and nuclei of the cells of these groups. Treatment of cells with SN50 [20 uM] or YC-1 [100 uM] resulted in a significant and almost complete inhibition of NFκB/p65 nuclear translocation. Images are representatives of three independent experiments. Scale bars, 200 mm.