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. 2013 Aug 26;8(8):e73862. doi: 10.1371/journal.pone.0073862

Figure 4. Identification of the target antigen by MALDI-TOF/TOF Mass analysis.

Figure 4

A. CBB-staining and Western blotting analysis with pancreas tissue sample. Band corresponding to Western blotting was cut from CBB-stained gel and analyzed by MALDI-TOF/TOF Mass. M: marker. B. Number of hits of candidate protein by MALDI-TOF/TOF Mass analysis and its significance score. Because the left part was lower than the threshold of protein score, that part had a lower confidence in candidate protein. Only Pdia2 had the high score, to be a sufficient candidate molecule as a target antigen. C. Sequence of Pdia2 identified by MALDI-TOF/TOF analysis and other four peptide sequences suggested from the analysis data. Sequence coverage was 12%. D. Pdia2 protein expression in BL21. M: marker, S: supernatant, P: pellet. E. Absorption test for WT mouse pancreas and stomach in 12-wk-old Aire KO or WT mice pooled sera. M: marker. F. The difference of the absorbance in WT or KO mouse CD4+ T cells stimulated with pdia2-pulsed BMDC. Each graph represents the mean ± SD of 3 replicates.