(A–C) Time-course analysis of neural progenitors marker PAX6 (A), neuronal marker MAP2 (B), glial marker GFAP (C) using real-time qPCR in wild-type and KNL1c.6125g > A neural spheroids.
(D) Immunostaining using antibodies against PAX6/NESTIN at 2 weeks, S100β/TUJ1 at 4 and 6 weeks, and SOX9/NEUN at 10 weeks in wild-type and KNL1c.6125G > A neural spheroids sections (scale bar, 200 μm).
(E) Wild-type and KNL1c.6125G > A neural spheroids at 10 weeks (scale bar, 500 μm).
(F) Area measurement of wild-type and KNL1c.6125G > A neural spheroids in a time course of 10 weeks.
One non-targeted wild-type clone, two wild-type clones, and three patient mutation clones derived from the same CRISPR-Cas9 targeting are plotted in each graph. Results are mean ± SEM. *p < 0.05, ***p < 0.001, ****p < 0.0001.