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. Author manuscript; available in PMC: 2016 Jul 16.
Published in final edited form as: Cell. 2015 Jul 16;162(2):375–390. doi: 10.1016/j.cell.2015.06.034

Figure 3. Decreased cell cycle length and increased neuronal/synaptic differentiation in ASD probands.

Figure 3

A, B, Cell cycle time determined by the formula Tc= Ts/(%BrdU/Ki67), where Tc= cell cycle time and Ts= S phase time. A, Representative image of double immunostaining for BrdU (red) and Ki67 (green) of undifferentiated iPSCs from a control individual and (B) early neuronal progenitors in monolayer cultures at TD11. *p < 0.05, ANOVA with family as covariant. To prepare monolayer cultures for neuronal progenitors the neural rosettes were dissociated into single cells and cultured in adhesion as monolayers until TD11. C, D, Representative images (C) and stereological quantification (D) of the proportion of proliferating Ki67+ neuronal progenitor cells in both control and ASD-derived organoids at TD31. E-J, Increased neuronal maturation and synaptic formation in ASD-derived neurons (TD 31). Representative images of controls and probands organoids at TD 31 labeled with MAP2 and SynI (E), or MAP2 and VGAT (H), and relative quantification of MAP2 density (F), number of SynI+ puncta (G), number of VGAT+ puncta (I), and number of VGLUT1+ puncta (J). The data in (F, G, I, J) are presented as means ± s.e.m; **p < 0.01, ***p< 0.001; t test analysis. Scale bars, 10 μm (A), 10–20 μm (B, as indicated), 10 μm (C), 5 μm (E, H).