Fig. 2
Cell cycle kinetics and spheroid cell death assay. A Cell cycle analysis in AIRE WT and AIRE- / - spheroids was performed by flow cytometry using PI staining; cell proportions were determined for each phase of the cell cycle following 12 and 24 h of spheroid formation. Distribution of cells at different cell cycle and Sub-G1 phases following 12 and 24 h of spheroid formation (N = 3). Values are expressed as mean ± SEM from data obtained in three independent experiments. Statistical analysis: unpaired t test. ns = no significant difference; 12 h: GO/G1 = **p = 0,0011; S = ns; G2M = ns; SubG1 = ***p = 0,0002. 24 h: G0/G1 = **p = 0,0026; S = ns; G2M = *p = 0,0105; SubG1 = ****p = < 0,0001 respectively indicate statistically significant differences when compared to AIRE WT spheroids. B The cell death analysis was measured using the eBioscience™ Annexin V Apoptosis Detection Kit (Invitrogen™), which determines the number of viable cells and detects cell death by apoptosis or necrosis. Living cells are shown in the lower left part of the respective plots; dying/dead cells shift to the right and upper part of the plot (representative plots from one experiment). The graphics show the total numbers (%) of apoptotic and necrotic cells (mean values ± SEM from three experiments). Statistical analysis: unpaired t test. *p = 0,0240,**p = 0,0023; ns = no significant difference; respectively, indicate statistically significant differences for the comparison between AIRE- / - and AIREWT spheroids