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. 2020 Aug 9;21(16):5713. doi: 10.3390/ijms21165713

Figure 2.

Figure 2

Expression of the peripheral tissue antigen (PTA) driving transcription factors autoimmune regulator (AIRE) and deformed epidermal autoregulatory factor 1 (DEAF1) in human LNSCs. (A) Expression of DEAF1 in cultured LNSCs of passage 2 was assessed by qPCR and compared between different donor groups (healthy individuals n = 5, RA-risk individuals n = 12 and RA patients n = 14). Relative quantity is displayed as median and interquartile range. (B) Intracellular expression of AIRE protein in cultured LNSCs was measured by flow cytometry. Histograms presenting % of positive cells in comparison to isotype staining. (C) The scatter plot represents the mean fluorescence intensity (MFI) of intracellular AIRE expression in cultured human LNSCs (passages 3–5) from individuals in different donor groups (healthy individuals n = 8, RA-risk individuals n = 9 and RA patients n = 4). Relative quantity is presented as median and interquartile range. (D) Representative pictures of immunofluorescence staining combined with confocal microscopy displaying AIRE (green) and nucleus (blue) in LNSCs (RA patient; passage 3) cultured on chambers slides. Isotype controls were negative. (E) Western blot analysis of AIRE protein expression in cultured LNSCs of 12 donors (passages 4–8) is shown. Actin was used as loading control and thymic tissue was used as positive control for AIRE expression.