Figure 3

Western blot of Synd1 shed from NMuMG cells by B. anthracis proteins in presence of 1% FCS for 24 h. Synd1 from culture supernatants was partially purified using DEAE ion exchange chromatography and analyzed before (lanes 1 to 4) and after (lanes 5 to 8) partial digestion for 5 h with both heparanase II (10 mU/ml) and chondroitin sulfate ABC lyase (20 mU/ml) to remove glycosaminoglycan chains. For digestion the amount of total protein in each reaction was adjusted approximately equal. Gel lanes correspond to untreated cells (1, 5) and cells challenged with: 0.1 μg/ml of AnlO for 4 h (2, 6); 1 μg/ml of AnlB for 4 h (3, 7); 1 μg/ml of LT for 24 h (4, 8). Synd1 core protein appears as a band of approximately 80 kD (open arrow).