Fig. 5.

Notch activation regulates Pou3f1 expression in ESC-derived MNs. (A-F) GFP, Lhx3 and Pou3f1 staining in day 6 EBs derived from parental H14IG#E3 ESCs and iNERT ESCs (without and with DOX/4HT induction). (G) Percentages of Pou3f1⁺ Lhx3^- (PN-like, red), Pou3f1^- Lhx3^-- (HMC-like, light blue), Pou3f1^- Lhx3⁺ (dark blue) and Pou3f1⁺ Lhx3⁺ (purple) cells among ESC-derived GFP⁺ MNs in day 6 EBs. The EBs were derived from ESC clones H14IG#E3 and iNERT without DOX/4HT induction. (H-M) GFP, Pou3f1 and Isl1/2 labelling in day 6 EBs. EBs were either differentiated according to the standard protocol (H,I) or treated the γ-secretase inhibitors LY411575 (J,K) or DAPT (L,M) from day 2 onwards. (N) Percentages of Pou3f1⁺ MNs among all GFP⁺ MNs in the absence or presence of γ-secretase inhibitor. Error bars indicate mean values from three independent experiments ± s.e.m. (G,N). ^*P<0.05 (paired Student’s t-test). Scale bars: 50 μm.