Fig. 3: Lack of CUL4B results in unbalanced differentiation into deep-layer and upper-layer neurons.

A Immunostaining of TUJ1, MAP2, TBR1 and CTIP2 on day 35 of neural differentiation. Scale bar, 50 μm. B Quantification of the percentage of TUJ1⁺ cells in all cells. N = 12. Quantifications of the percentage of TBR1⁺ cells (C) or CTIP2⁺ cells (D) in TUJ1⁺ cells. N = 12-14. E Quantification of the percentage of MAP2⁺ cells in all cells. N = 12. F mRNA levels of MAP2, TBR1, CTIP2 and FOXP2 on day 35. G Immunostaining of MAP2, SATB2, CUX1 on day 60 of neural differentiation. Scale bar, 50 μm. Quantifications of SATB2⁺ cells (H) and CUX1⁺ cells (I). N = 10. J mRNA levels of CUX1, SATB2 on day 60. K Representative images of day 60 organoids stained with antibodies against PAX6 and SATB2. Scale bar, 50μm. The bar graph shows the proportion of SATB2⁺ cells in the isogenic control (Correct) and the patient-derived (Patient) organoids. N = 12. Data are presented as the mean ± SEM. The statistical significance was determined using one-way ANOVA with Tukey test for comparing three or more samples and two-tailed unpaired t-test for two-sample comparison. *: P < 0.05; **: P < 0.01; ***: P < 0.001. ns: no significance.