Anti-ZnT8 immunoreactivity of human sera.
A, purification of whole IgG from human ZnT8A-positive and ZnT8A-negative sera. B, formation of ZnTA-ZnT8·GFP complexes as revealed by GFP fluorescence size exclusion HPLC of detergent-solubilized ZnT8·GFP exposed to purified IgG from diabetic patients as indicated. IgG from non-diabetic control subjects was analyzed in parallel as a negative control. C, representative confocal images of immunofluorescence staining of WT or ZnT8 KO cells using IgG from diabetic patients or non-diabetic control subjected as indicated. Scale bar, 20 μm. D, histograms of surface immunofluorescence intensities measured in >10,000 WT or ZnT8 KO INS-1E cells that were stained by IgG from diabetic patients or non-diabetic control subjects as indicated. Solid lines are fits of histograms to a Lorentzian distribution.