Fig. 6. APS-1 mutations C302Y and C311Y increase PML association.

a 3D structure of Aire PHD1 domain bound to histone H3-derived peptide (PDB ID: 2KFT [10.2210/pdb2KFT/pdb]). C302 and C311, which coordinate Zn²⁺ ions, are mutated in APS-1 patients (highlighted in orange). b Transcriptional activity of hAire WT, C302Y and C311Y. Experiments were performed as in Fig. 1g and are presented as mean ± s.d., n = 3. c Representative fluorescence microscopy images of hAire C302Y and C311Y variants in 4D6 cells. Right, quantitation of Aire foci colocalized with PML bodies. n = number of Aire foci examined per sample. Statistical significance comparisons were calculated using a two-tailed Student’s t-test for two population proportions where each population consists of all individual Aire foci examined. ***p = 2.6e-18 (C302Y) and 0.0004 (C311Y). d SUMOylation analysis of hAire WT, C302Y (top) and C311Y (bottom). Experiments were performed as in Fig. 3c. e Chromatin fractionation analysis of hAire WT, C302Y and C311Y. Experiments were performed as in Fig. 3d.