(A) Proliferation of NSPCs from E14 nestin-Cre cKO embryos was retarded in primary culture but rescued by transfection of Apc4 (n = 4). **p<0.01 in one way ANOVA with post hoc Tukey’s test.
(B) FACS analysis showed G2/M accumulation and a mild increase in the G1 population in NSPCs derived from E14 cKO embryos. Apc4 transfection rescued delayed proliferation and prevented cyclin B accumulation in NSPCs from cKO embryos.
(C) The pial-to-apical surface area ratio reflects the proliferation of in-utero transfected EGFP-positive cells after a defined time period. pApc4-IRES-hrGFPII or pIRES-hrGFPII were electroporated into the ventricular zone of E13 embryos (nestin-Cre control and cKO mice), and the brains were analyzed at E18.
(D) Pial-to-apical ratio was deduced from 3D reconstruction of the rostral-to-caudal axis serial sections of six embryos in each genotype. The ratio was decreased in Pqbp1-cKO embryos, reflecting the decreased cell cycle times of NSPCs, but it was rescued by Apc4 expression. **p<0.01 or *p<0.05 in one way ANOVA with post hoc Tukey’s test.