1. Transitional embryoid-body-like cells (Fujimori 140 et al., 2017) were seeded at a density of 10 cells;( Hoashi;therefore, we generated 137 neurons with the characteristics of this brain region. Human iPSCs were pretreated for 5 days 138 with 3 ?M SB431542,2015
2. Japan) with specifically selected growth factors and inhibitors at 4% O 2 and 5% 142 CO 2 ; growth factors and inhibitors included 1� B-27 supplement (Thermo Fisher Scientific), 143 0.5% penicillin/streptomycin, 20 ng/mL FGF-2, 2 ?M SB431542, and 3 ?M CHIR99021. The 144 neurosphere culture was started on days in vitro (DIV) 0; neurospheres were passaged at a 145 density of 50 cells/?L on DIV7 and DIV14. The neurosphere culture medium contained the 146 following additives;Saitama Bio;) on DIV1-21. On DIV21, neurospheres 149 were replated on dishes coated with a poly-L-lysine solution and fibronectin and
3. 20 ng/mL glial cell-derived neurotrophic factor 155 (GDNF; Alomone Labs;San Bdnf; Biolegend;Fisher Scientific), 0.5% penicillin/streptomycin, 20 ng/mL brain-derived neurotrophic factor 154
4. Figure 5—figure supplement 4. Snf7 activation allows ESCRT-III helix formation.