Figure 4.

Differential Effects of NF1 Mutations on Cerebral Organoid Progenitor Cell Dynamics and Neurogenesis

(A and B) SOX2⁺ NPCs in the ventricular zones of group 1 NF1-mutant cerebral organoids exhibit (A) 1.3- to 1.4-fold increased proliferation (Ki67⁺ cells; white arrowheads) and (B) 2- to 3-fold increased cell apoptosis (cleaved caspase-3; white arrowheads) compared with control and group 2 cerebral organoids at 16 DIV.

(C–E) Decreased numbers of (C and D) early immature neurons (NeuroD1; TUJ1 white arrowheads) and (E) late immature neurons (MAP2; white arrowheads) migrating into the periventricular zone of group 2 compared with group 1 and control cerebral organoids at 16 DIV.

(F) Quantifications of %Ki67⁺ NPCs, %cleaved caspase-3⁺ NPCs and NeuroD1⁺ immature neurons in NF1-mutant cerebral organoids compared with controls at 16 DIV.

Each dot represents an independently generated data point derived from separate experiments and the two different clones for each line are denoted as black versus gray dots. All data are represented as means ± SEM. One-way ANOVA with Dunnett post-test. n.s., not significant. Scale bars, 10 μm (C) and 50 μm (A, B, D, and E).