bRGC-driven direct neurogenesis, quantification of PTPRZ+ bRGCS.

(A) Immunostaining showing PAX6, TBR2, and the bRGC marker PTPRZ in week 6 organoids. Scale bar = 50 μM (B) Quantification of bRGCs using PTPRZ+TBR2− bRGCs. (C) Schematic showing the experimental design of the BrdU treatment at week 6. Created with BioRender.com. (D) Immunostaining showing CTIP2, HOPX, BrdU, and DAPI in week 6 organoids. Scale bar = 50 μM (E) Quantification of neurogenic bRGCs, which are CTIP2+ cells located next to HOPX+ cells. These cells may represent neurons born directly from bRGC to EN. For all quantifications, n = 3–8 organoids per sample per genotype was used. In each organoid, 3–9 cortical structures with clear lamination patterns were examined. Data are represented in bar graphs as mean ± STD with individual data as dots; n.s. = not statistically significant, *p < 0.05, Kruskal–Wallis ANOVA test with Dunn’s multiple comparisons test as a post hoc. The numerical values that were used to generate the graphs can be found in S1 Data. BrdU, 5-bromo-2-deoxyuridine; bRGC, basal radial glial cell; EN, excitatory neuron.

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