Angiogenic inhibitors activate NF-κB in endothelial precursor cells. (A-B) NF-κB nuclear localization in bone marrow endothelial progenitor cells (EPCs) stimulated to differentiate for 7 days with VEGF (10 ng/mL) and treated with TSP1 (100nM) or PEDF (10nM) was visualized by immunostaining for p65 (A) and measured by Western blot analysis of nuclear extracts for p65 (top) and p50 (middle; B). Loading was assessed by TBP (bottom). Three independent experiments were performed with similar results. (C) EPC colony formation with and without PEDF (10nM). Images were viewed with a Nikon Eclipse TE 2000-U microscope with a 40×/0.60 air objective and Fluoromount G imaging medium (Southern Biotech), and captured with a Nikon LH-M100 C-1 camera. Metamorph, Adobe Photoshop CS3 Extended Version 10.01, and Corel Photo-Paint 10 software were used.