REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Mouse monoclonal anti-alpha-Tubulin (B-5-1-2)	Thermo Fisher Scientific	Cat#32-2500; RRID: AB_2533071
Rabbit monoclonal anti-GSK-3α (D80E6)	Cell Signaling Technology	Cat#4337; RRID: AB_10859910
Mouse monoclonal anti-GSK-3 beta (clone 7)	BD Biosciences	Cat#610201; RRID: AB_397600
Mouse monoclonal anti-β-catenin (clone 14)	BD Biosciences	Cat#610153; RRID: AB_397554
Rabbit polyclonal anti-phospho-β-catenin (Ser33/37/Thr41)	Cell Signaling Technology	Cat#9561; RRID: AB_331729
Rabbit monoclonal anti-GAPDH (D16H11)	Cell Signaling Technology	Cat#5174; RRID: AB_10622025
Rabbit monoclonal anti-Axin1 (C76H11)	Cell Signaling Technology	Cat#2087; RRID: AB_2274550
Mouse monoclonal anti-Oct-3/4 (C-10)	Santa Cruz Biotechnology	Cat#sc-5279; RRID: AB_628051
Goat polyclonal anti-Sox-2 (Y-17)	Santa Cruz Biotechnology	Cat#sc-17320; RRID: AB_2286684
Goat polyclonal anti-mouse Nanog	R&D Systems	Cat#AF2729; RRID: AB_2150103
Mouse monoclonal anti-Neuronal Class III β-Tubulin (TUJ1)	Covance Research Products Inc	Cat#MMS-435P; RRID: AB_2313773
Mouse monoclonal anti-Myosin (MF-20)	Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA	Cat#MF-20; RRID: AB_2147781
Mouse monoclonal IgG1 Isotype Control	Cell Signaling Technology	Cat#5415S; RRID: AB_10829607
Chemicals, Peptides, and Recombinant Proteins
CHIR99021	Division of Signal Transduction Therapy, University of Dundee, UK	CAS: 252917-06-9
PD0325901	Division of Signal Transduction Therapy, University of Dundee, UK	CAS: 391210-10-9
N-2 Supplement (100X)	Thermo Fisher Scientific	Cat#17502048
B-27™ Supplement (50X), serum free	Thermo Fisher Scientific	Cat#17504044
3MB-PP1	Cayman Chemical	Cat#17860; CAS: 956025-83-5
1NA-PP1	Cayman Chemical	Cat#10954; CAS: 221243-82-9
1NM-PP1	Cayman Chemical	Cat#13330; CAS: 221244-14-0
Leukemia inhibitory factor	Our own lab	N/A
ATP, [γ-32P]- 3000Ci/mmol 5mCi/ml EasyTide, 250 μCi	PerkinElmer	Cat#BLU502H500UC
Peptide substrate (RRRAAEELDSRAG{pSer}PQL)	GenScript	N/A
Critical Commercial Assays
Leukocyte Alkaline Phosphatase Kit	Sigma-Aldrich	Cat#85L3R
Dual-Luciferase Reporter Assay System	Promega	Cat#E1910
iTaq™ Universal SYBR® Green Supermix	Bio-Rad	Cat#1725124
RNeasy Mini Kit	Qiagen	Cat#74104
Deposited Data
RNA-seq data of gene expression profiles in mouse ES cells upon GSK3α or GSK3β inhibition	This paper	GEO: GSE98080
Full western blots	This paper	Mendeley Data: https://data.mendeley.com/datasets/sncsy3phsy/draft?a=87bb5918-ac94-46e4-a46b-40dac003f5ef
Experimental Models: Cell Lines
Mouse ES-E14 cell line	Ying et al., 2008	RRID: CVCL_C320
Mouse ES-46C cell line	Ying et al., 2003	RRID: CVCL_Y482
Gsk3α−/− mouse ES cells	This paper	N/A
Gsk3β−/− mouse ES cells	This paper	N/A
Gsk3α−/−; Gsk3β−/− mouse ES cells	This paper	N/A
Oligonucleotides
Gapdh: F-tgtgagggagatgctcagtg, R-tgttcctacccccaatgtgt	Qiu et al., 2015	N/A
Esrrb: F-aacagcccctacctgaacct, R-ctcatctggtccccaagtgt	Tai et al., 2014	N/A
Tfcp2l1: F-aggtgctgacctcctgaaga, R-gttttgctccagctcctgac	Qiu et al., 2015	N/A
Oct4: F-gaagcagaagaggatcaccttg, R-ttcttaaggctgagctgcaag	Tai et al., 2014	N/A
Rex1: F-tcactgtgctgcctccaagt, R-gggcactgatccgcaaac	Qiu et al., 2015	N/A
Klf2: F-aggcctgtgggttcgctataaa, R-ggcaaattatggctcaaagtagcag	Qiu et al., 2015	N/A
Gsk3α sgRNA. 5′-TGATTGGTAATGGCTCATT-3′	This paper	N/A
Gsk3β sgRNA. 5′-ATTCCAAAGGAATGGATAT-3′	This paper	N/A
Recombinant DNA
PiggyBac vector (CAG promoter)	Ye et al., 2016	N/A
px330	Cong et al., 2013	Addgene Plasmid #42230
px330-P2A-puro	This paper	N/A
pGL2-SuperTop plasmid	Kim et al., 2013	N/A
pRL-TK	Promega	Cat#E2241
pET-41a	EMD Biosciences	Cat#70556-3
Software and Algorithms
FIJI (ImageJ)	NIH	ImageJ ver2
Excel	Microsoft	2016
Prism	GraphPad	Ver 5
Partek Flow	Partek Inc.	V6.0.17.0219