Because IgG titers in TGC-induced EAO mice were higher than those of IgA and IgM, IgG was used as a secondary antibody for detecting TGC-specific AIs. Spot numbers were only assigned to spots identified by mass spectrometry (MS) using TGC autoantibody as a primary antibody (Table S1). All spots from EAO and control were detected by MS. However, because the spots in which multiple proteins are mixed together, they cannot be identified by MS, thus spot number was not attached. All proteins that visualized on a silver-stained gel were excised and processed for protein identification using MS (Fig. 3a,d). Altogether, isoelectric point (pI) 4–7 set of 29 protein spots (Fig. 3c and Table S1) and pI 6–9 set of 10 proteins spots (Fig. 3f and Table S1) were identified that reacted with EAO serum sample, whereas pI 4–7 set of 23 protein spots (Fig. 3b and Table S1) and pI 6–9 set of two proteins spots (Fig. 3e and Table S1) were identified that reacted with control sera. Some spots (spot no. 1, 3–9, 11–14, 17–20, 22–23, 25–26, 28–31, and 33) overlapped between the EAO and control serum. Other spots (spot no. 2, 10, 15, 16, 21, 24, 27, 32, and 34–36) in EAO serum were not identified in control sera. In effect, we have identified the spots which reacted only with the serum IgG of EAO as TGC-specific AIs. These spots were summarized in Table 1.