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. 2020 Apr 2;11:1625. doi: 10.1038/s41467-020-15448-w

Fig. 5. Isolated Aire CARD multimers associate with PML bodies.

Fig. 5

a Representative fluorescence microscopy images (3 independent experiments) of mAire CARD fused with monomeric GFP (CARD-mGFP) transiently expressed in 4D6 cells. Note that mAire residues 1–174 containing both Aire CARD and nuclear localization signal (NLS) were used in the fusion construct. GFP fluorescence and anti-PML staining were used for imaging CARD-mGFP and PML bodies, respectively. b Chromatin fractionation analysis of NLS-mGFP (mAire aa 105–174 fused with mGFP) and CARD-mGFP. Experiments were performed as in Fig. 3d. c Representative fluorescence microscopy images (two independent experiments of CARD-mGFP and HA-tagged mAire upon their co-expression in 4D6 cells. GFP fluorescence was used for imaging CARD-mGFP and immunostained with anti-HA and anti-PML. d Transcriptional activity of mAire (black circle) and its changes upon co-expression with CARD-mGFP (red circle) in 293 T cells. Each circle represents 0.6 μg/ml DNA transfected. Experiments were performed as in Fig. 1g and presented as mean ± s.d., n = 3. P-values (two-tailed t-test) were calculated in comparison to WT mAire. *p < 0.05; p > 0.05 is not significant (ns). Exact p-values are provided in the Source Data File.