(A) Effect of VEGF infusion on NF-κB binding to DNA. Mice were treated with VEGF or PBS for 7 or 28 days. BM cells were obtained and lin⁻ progenitor cells were prepared as described in Materials and Methods. Cells were stimulated with 10 ng/mL murine TNF-, nucleoproteins were extracted, and EMSA was performed as described in Materials and Methods. Control, BM cells from mice treated with PBS for 7 days; VEGF, mice treated with VEGF (50 ng/h) for 7 or 28 days; M, binding of nucleoproteins from control TNF-–stimulated BM cells to mutant DNA sequence (negative control); 1, nonstimulated cells; 2, cells stimulated with 10 ng/mL TNF- for 10 minutes; 3, cells stimulated with TNF- for 20 minutes. Three experiments with the same results were performed. (B) Effect of tumor cells on NF-κB binding activity in BM cells. Mice were injected subcutaneously with 2 × 10⁵ D459 tumor cells. Mice were killed on day 18 (tumor size, 80 mm²) or on day 32 (tumor size, 300 mm²) after tumor injection. Lin⁺ cells were removed and EMSA was performed exactly as described in the legend to Fig 10A. Control, tumor-free mice. 1, 2, 3: The same as in Fig 10A. Two independent experiments with the same results were performed.