Figure 4.

NEUROG1 Overexpression in Proliferating Progenitors

(A) Automated nuclear counts with Hoechst-labeled nuclei are marked within green circles.

(B) Doubling times for PB-T-EGFP and PB-T-Neurog1 cells cultured in the absence or presence of 1 μg/mL Dox (n = 5 for each condition).

(C–F) Cells were fixed and labeled with Hoechst and subjected to EdU detection in (C) PB-T-EGFP – Dox (n = 3), (D) PB-T-EGFP + Dox (n = 3), (E) PB-T-Neurog1 – Dox (n = 3), (F) PB-T-Neurog1 + Dox (n = 3) cultures. Scale bars, 20 μm.

(G) Relative percentage of cells for PB-T-EGFP and PB-T-Neurog1 cells cultured in the absence or presence of 1 μg/mL Dox (n = 3 for each condition), and percentage of EdU-labeled cells (n = 3 for each condition).

(H) Transcript levels of NeuroD1 and Cdk2 from PB-T-EGFP and PB-T-Neurog1 cells cultured in the absence or presence of 1 μg/mL Dox (n = 3 for each condition).

(I) Western blot of CDK2 in proliferating PB-T-EGFP (n = 3) and PB-T-Neurog1 (n = 3) cultured in 1 μg/mL Dox.

(J) Quantification of CDK2 protein levels from western blots (n = 3).

Error bars denote ±SEM. ns, not significant; ^∗∗p < 0.01, ^∗∗∗p < 0.001.