Figure S3.

NOTCH2NLB Promotes Cell-Cycle Re-entry and Maintenance pf Cortical Progenitors, Related to Figure 5

(A–C) The percentage of mitotic cells and the cells in G2/S phase are quantified using anti-phospho Histone H3 and EdU labeling Cell cycle exit in the human cortical progenitors derived from ESC; NOTCH2NLB introduced by lentiviral infection at day 30 of differentiation, EdU incorporated 24 hours before fixation at day 7 of overexpression.

(F–S) In utero electroporation of NOTCH2NLB full length (N2NL-FL), NOTCH2NL-EGF repeats deletion (ΔEGF) and mouse NOTCH1 intracellular domain (NICD) in E13.5 mouse cortex, followed by analysis at E15.5. Bin analysis of fractions of mCherry+ electroporated cells in four regions, the CP, IZ, SVZ and VZ (F and G) PAX6 immunoreactivility was examined to quantify the proportion of apical and basal RG progenitors following NOTCH2NL overexpression (H-M). Proportions of PAX6-positive cells in the VZ (L) and SVZ (M) among all electroporated cells in whole cortical thickness are quantified. TBR2 immunoreactivility is examined to quantify the proportion of basal/intermediate progenitors in these four conditions (N-S). Proportion of TBR2-positive cells in the VZ (R) and SVZ (S) among all electroporated cells in whole cortical thickness are quantified.

Data are represented as mean ± sem and p values by Student’s t test (B, C and E) one-way ANOVA and bonferroni post hoc test (H, I, P, Q, X and Y). Scale bars; 100μm (A, D, F, L, R and T) and 20μm (A1, A2, D1, D2, J, K and S).