Immunoprecipitation (IP) of MFG-E8 was done as previously described (41 (link)-42 (link)). Wound-edge tissue lysates (100µg) of diabetic animals were incubated with 5 μg of MFG-E8 mAb (clone 2422) overnight in a rotisserie shaker at 4°C. Protein A-agarose beads (Invitrogen) were prewashed with lysis buffer (150 mm KCl, 25 mm Tris-HCl, 5 mm EDTA, 0.5% IgePal, 1 mm PMSF, protease inhibitor) and incubated with the lysate-antibody mix for 3h at 4°C to immunoprecipitate MFG-E8. The beads were then washed three times with ice-cold lysis buffer and the immunoprecipitated complexes were washed four times with lysis buffer. For Western blot, the IP samples were subjected to SDS/PAGE after reduction with 1M DTT as previously described (37 (link), 41 (link)) and probed with anti-AGE antibody (1: 1000 dilution, Abcam). The membrane was stripped and reprobed with anti-MFG-E8 antibody (Clone 18A2-G10).