Figure 3.

Analysis of hESC differentiation and neurogenesis at day 21

qPCR analysis of SREBF1 (A) and ABCA1 (B) at day 21. ^∗∗∗ p < 0.001 versus GW3965(−) condition. ††† p < 0.001 versus day 16 (n = 6 independent experiments).

(C) Violin plots of SOX2 and DCX generated from scRNA-seq data of developing human ventral midbrain.

(D) Immunostaining of SOX2⁺;DCX⁺ cells at day 21. Scale bar, 100 μm.

(E and F) Quantification of SOX2⁺ cells (E) and DCX⁺ cells (F). ^∗∗p < 0.01 and ^∗∗∗p < 0.001 versus GW3965 (−) condition (n = 8 independent experiments).

(G) Immunostaining of SOX2⁺;DCX⁺ cells at day 16 and EdU⁺; DCX⁺ cells at day 21, after GW3965 treatment (day 16–21). EdU pulse was performed for 4 h at day 16 and EdU detection was performed at day 21. Scale bar, 100 μm.

(H) Quantification of EdU⁺;DCX⁺/DAPI⁺ cells at day 21. ^∗∗p < 0.01 versus GW3965(−) condition (n = 6 independent experiments).

(I) Immunostaining of LMX1A⁺ and NGN2⁺ cells at day 21. Scale bar, 100 μm.