Figure 4. Proliferation studies of day-30 cerebral organoids.

(A–D) Immunofluorescence of Ki67⁺, EdU⁺, SOX2⁺, and PAX6⁺ proliferating radial glial progenitors, mature MAP2⁺, TUJ1⁺ neurons, and deep-layer VI TBR1⁺ excitatory neurons after 30 days of differentiation. Scale bar: 20 μm. (E–H) Quantification of the proportion of Ki67⁺, SOX2⁺, PAX6⁺, and EdU⁺ cells in trisomy 21 and euploid organoids after 30 days of differentiation. n = 15–42 VZ-like regions in at least 5 organoids per cell line. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test.