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 |