REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
NKX2.1 (1:100)	Abcam	ab76013
FOXA2 (1:200)	Cell Signaling Technology	8186T
SOX17 (1:200)	Thermo Fisher	AF1924-SP
SOX2 (1:200)	Cell Signaling Technology	2748
SOX9 (1:500)	Abcam	ab185230
ZO1 (1:200)	Thermo Fisher	40-2200
SYP (1:500)	Abcam	ab32127
SYP (1:100)	Santa Cruz Biotechnology	sc-17750: SYP (D-4)
ENO2 (1:100)	Santa Cruz Biotechnology	sc-271384: Enolase (A-5)
CHGA (1:100)	Proteintech	60135-1-Ig
ROBO2 (1:100)	R&D Systems	AF3147
PGP9.5 (1:100)	Santa Cruz Biotechnology	sc-271639: UCH-L1 (C-4)
EPCAM (1:500)	R&D Systems	AF960
VIM (1:500)	Sigma-Aldrich	V6630
Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488	Thermo Fisher Scientific	A-21206
Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555	Thermo Fisher Scientific	A-31572
Donkey anti-rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 594	Thermo Fisher Scientific	R37119
Donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 555	Thermo Fisher Scientific	A-21432
Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488	Thermo Fisher Scientific	A-21202
Donkey anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 555	Thermo Fisher Scientific	A-31570
Chemicals, Peptides, and Recombinant Proteins
mTeSR1 media	STEMCELL Technologies	85850
Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F-12)	Life Technologies	11330032
Roswell Park Memorial Institute (RPMI) 1640 Medium	Life Technologies	11875-093
FBS	Genesee Scientific	25-514
Geltrex™ LDEV-Free Reduced Growth Factor Basement Membrane Matrix	Thermo Fisher Scientific	A1413202
ReLeSR	STEMCELL Technologies	05782
Accutase	STEMCELL Technologies	7920
Y-27632 (Rock inhibitor)	Selleckchem	S1049
SB431542	Selleckchem	S1067
Recombinant Human FGF7/KGF	Peprotech	100-19
Recombinant Human FGF2	Peprotech	100-28
Recombinant Human FGF10	Peprotech	100-26
Activin A	Humanzyme	HZ-1140
Recombinant Human Wnt-3α	R&D Systems	5036-WN-010
Recombinant Human Noggin	Peprotech	120-10C
Recombinant Human BMP4	Humanzyme	HZ-1045
Bovine Serum Albumin (BSA)	Sigma-Aldrich	A2058
Phosphate-buffered saline (PBS), 1×	Genesee Scientific	25-507
DAPT	Cayman Chemical	208255-80-5
Laminin	Life Technologies	23017015
Fibronectin	R&D Systems	1918-FN-02M
Collagen IV	Sigma-Aldrich	C7521-5MG
Iscove's Modified Dulbecco's Media (IMDM)	Life Technologies	12440053
Ham’s F12 Media	Life Technologies	11765-054
B-27 with RA supplement (50×)	Life Technologies	17504-044
N-2 supplement (100×)	Life Technologies	17502-048
Glutamax	Life Technologies	35050
Ascorbic Acid	Sigma-Aldrich	A8960-25G
α-Monothioglycerol (MTG)	Sigma-Aldrich	M1753-100ML
Bovine Brain Extract	Lonza	CC-4098
Epinephrine	Sigma-Aldrich	E4642
Ethanolamine	Sigma-Aldrich	E0135
Fetal Clone II	Hyclone	SH30066
Hydrocortisone	Sigma-Aldrich	H4001
Insulin	Sigma-Aldrich	I9278
Phosphoryl ethanolamine	Sigma-Aldrich	P0503
Retinoic Acid	Sigma-Aldrich	R2625
3,3′,5-Triiodo-L-thyronine sodium salt	Sigma-Aldrich	T6397
Transferrin	Sigma-Aldrich	T8158
Ultroser G serum substitute	Crescent chemicals	67042.1/S
Paraformaldehyde (PFA)	VWR	100496-496
CAS Block	Thermo Fisher Scientific	008120
Triton X-100	Sigma-Aldrich	T8787
Phosphate Buffered Saline (PBS)	Genesee Scientific	25-507
Immunomount	Thermo Fisher Scientific	9990402
Vectashield with DAPI	Vector Laboratories	H-1500
DAPI	Sigma-Aldrich	D9542
DNase	Worthington Biochemical Corporation	LS003172
SYBR Green	Bio-rad	1725125
Critical Commercial Assays
Qiagen RNA Easy Mini Kit	Qiagen	74004, 74134
Protoscript first strand synthesis kit	New England Biolabs	E6560S
Enolase Quantikine ELISA Kit	R&D	DENL20
Experimental Models: Cell Lines
Healthy human lymphoblastoid cell line-derived iPSCs (iPSC 1). Lymphoblastoid cell line reprogrammed by Ichida Lab USC	National Institute of Neurological Disorders and Stroke (NINDS) Biorepository, Coriell Institute	ND00184 (original lymphoblastoid cell line)
Healthy human lymphoblastoid cell-derived iPSCs (iPSC 2). Lymphoblastoid cell line reprogrammed by Ichida Lab USC	National Institute of Neurological Disorders and Stroke (NINDS) Biorepository, Coriell Institute	ND03719 (original lymphoblastoid cell line)
Human foreskin fibroblast-derived iPSC line (FiPSC #3C15)	Ryan Laboratory, USC	Derived from BJ fibroblasts (ATCC, CRL-2522)
Deposited Data
Raw and analyzed iPNEC data	Hor et al., 2020	GEO: GSE146990
Oligonucleotides (qPCR Primers)
Gene	Forward Primer	Reverse Primer
β-Actin	5′-CATGTACGTTGGTATCGAGGC-3′	5′-CTCCTTAATGTCACGCACGAT-3′
HES1	5′-ACGTGCGAGGGCGTTAATAC-3′	5′-GGGGTAGGTCATGGCATTGA-3′
HEY1	5′-AGAGTGCGGACGAGAATGGAAACT-3′	5′-CGTCGGCGCTTCTCAATTATTCCT-3′
ASCL1	5′-CCCAAGCAAGTCAAGCGACA-3′	5′-AAGCCGCTGAAGTTGAGCC-3′
GRP	5′-AAAGAGCACAGGGGAGTCTTC-3′	5′-TCCTTTGCTTCTATGAGACCCA-3′
DLL3	5′-CGTCCGTAGATTGGAATCGCC-3′	5′-TCCCGAGCGTAGATGGAAGG-3′
GAD67	5′-GCTTCCGGCTAAGAACGGT-3′	5′-TTGCGGACATAGTTGAGGAGT-3′
ENO2	5′-CTGATGCTGGAGTTGGATGG-3′	5′-CCATTGATCACGTTGAAGGC-3′
ROBO2	5′-GTTTGTGTTGCGAGGAACTATCT-3′	5′-GTTTTGTCGGAAGTCATCTCGTA-3′
SYP	5′-CTCAGCATCGAGGTCGAGTTC-3′	5′-GAGGAGTAGTCCCCAACTAAGAA-3′
SST	5′-CCCAGACTCCGTCAGTTTCT-3′	5′-AAGTACTTGGCCAGTTCCTGC-3′
SV2	5′-TAGACCAGGCACTCATTGGC-3′	5′-ACCCCTCCCCACAGTTACTT-3′
PENK	5′-TCTGAACCCGGCTTTTCCAA-3′	5′-TACGCAAGCCAGGAAGTTGAT-3′
Software and Algorithms
ImageJ	National Institutes of Health	https://imagej.nih.gov/ij/
Quantitative RT-PCR software	Thermo Fisher Scientific	7900HT SDS software v2.1
Other
6-Well Cell Culture Plates	Genesee Scientific	25-105
Transwell® Inserts, Sterile, Corning® (3470), 24 well plate	VWR	29442-082
Vacuum Filter Systems, 500 mL	Genesee Scientific	25-227
0.45 μm pore filter	Genesee Scientific	25-246
BD Disposable Syringes with Luer-Lok™ Tips (1 and 5 mL)	VWR	BD309646, BD328438