Figure 4

Autoantibodies against fractalkine are present in NOD mice. Tissue lysates prepared in phosphate-buffered saline supplemented with protease inhibitors were run on an SDS gel and transferred to an Immobilon membrane (Millipore, Billerica, MA, USA). Membranes were then incubated with purified IgG from either NOD (N) or Balb/c (Ba) mice. Binding of autoantibodies was detected by anti-mouse-HRP and electrochemical luminescence (ECL) analysis. Presence of specific fractalkine fragments in the lysate was detected using a polyclonal anti-fractalkine antibody (a-Fr). Numbers represent mass (kDa), as determined using a molecular weight marker.