Fig. 4

Spheroid morphology A Internal structure of the spheroids Aire^WT and Aire^−/− .Light microscopy (H&E staining) of histological analysis of spheroid growth from 12 to 48 h. In the upper part of the Figure is represented AIRE^WT spheroid at 12, 24,36 and 48 h of adhesion, showing regular round centers, exhibiting well-defined contours, whereas in the lower part of the Figure (AIRE^−/− spheroids), the cells present a lobule-like form, exhibiting ill-defined contours at 12 and 24 h of adhesion, resuming the regular round formation after 36 h of culture. B Area of spheroids during its growth from 12 to 48 h. The area was measured and compared through paired student t-test (p-value = *** 0.0007; *** 0.0008; * 0.0368; ns 0.9861 respectively). ns: no significant difference. C Scanning electron microscopy (SEM) of the spheroids Aire^WT and Aire^−/−. A representative SEM images comparing 24 h Aire^WT vs. Aire^−/− spheroids. Representative images from 20 spheroids analyzed for each time-point. Scanning electron microscope (Jeol JSM -6610 LV), 5 kV acceleration voltage