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Published in final edited form as: Nat Neurosci. 2023 Nov 9;26(12):2090–2103. doi: 10.1038/s41593-023-01477-3

Extended Data Figure 4 — A. Single rosette from the Patient^Corrected cell line expressing SOX2 and PAX6 markers.

B. Single rosette from the Patient^p.Q503X cell line expressing SOX2 and PAX6 markers.

C. Single rosette from the 03231^Control cell line expressing SOX2 and PAX6 markers.

D. A single rosette was generated from Patient^Corrected line. The rosette is composed of cells positive for the neural progenitor marker SOX2 and SYNGAP1. SYNGAP1 is also highly expressed at the apical wall of the lumen. The tight junction protein TJP1 labels the central luminal space of the rosette. Merged images show colocalization of DAPI, SYNGAP1, and TJP1.

E. A single rosette was generated from Patient^p.Q503X line. The rosette is composed of cells positive for the neural progenitor marker SOX2 and SYNGAP1. Merged images show colocalization of DAPI, SYNGAP1, and TJP1. The Patient^p.Q503X single rosettes display a larger and more irregularly shaped central luminal space.

F. A single rosette was generated from 03231^Control line. The rosette is composed of cells positive for the neural progenitor marker SOX2 and SYNGAP1. SYNGAP1 is also highly expressed at the apical wall of the lumen. Merged images show colocalization of DAPI, SYNGAP1, and TJP1.

G. Single rosette from the 03231^p.Q503X cell line expressing SOX2 and PAX6 markers.

H. Single rosette from the 03231^RGD cell line expressing SOX2 and PAX6 markers.

I. A single rosette was generated from 03231^p.Q503X line. The rosette is composed of cells positive for the neural progenitor marker SOX2 and SYNGAP1. Merged images show colocalization of DAPI, SYNGAP1, and TJP1. The tight junction protein TJP1 is weakly expressed with little to no central luminal organization.

J. A single rosette was generated from 03231^RGD line. The rosette is composed of cells positive for the neural progenitor marker SOX2 and SYNGAP1. Merged images show colocalization of DAPI, SYNGAP1, and TJP1. The tight junction protein TJP1 is weakly expressed with no central luminal organization.

K. A Survival curve for organoids generated from the 03231^RGD line. Data was collected from 10 independent differentiations, each representing an average of 6 organoids for each time point. Single dots represent total averages for that time point. Student’s t-test was performed (Day 30 P=0.0134, Day 60 P<0.0001). Data is shown as mean ± SEM.