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. 2015 Feb 2;5:8162. doi: 10.1038/srep08162

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(a) Human annexin A2 protein was overexpressed in E. coli BL21 after IPTG-induced expression. (b) The annexin A2 protein was purified using Ni-NTA resin. (c) Verification of annexin A2 using mass spectrometry. (d) Protein marker. (e) Immunoprecipitation was performed to determine whether annexin A2 was a real autoantigen of BD. Annexin A2 was clearly present in the immunoprecipitates. (f) Purified annexin A2 protein served as a control in immunoprecipitation analysis. (g) The EA.hy926 staining pattern of sera from BD patients in competitive immunofluorescence assays which were used to analyze the inhibitory activity of the annexin A2 protein. (h) Pre-incubation of serum with annexin A2 resulted in inhibition of the staining. (i) EA.hy926 cells incubated with HC serum served here as a control.