KEY RESOURCES TABLE
REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
CD140a-APC | eBioscience | Cat#17-1401-81; RRID:AB_529482 |
NG2-AF488 | Millipore | Cat#AB5320A4; RRID:AB_11203143 |
OLIG2 | Proteintech | Cat#13999-1-AP; RRID:AB_2157541 |
OLIG2 | Millipore | Cat#AB9610; RRID:AB_570666 |
OLIG2 | Millipore | Cat#MABN50; RRID:AB_10807410 |
MBP | Abcam | Cat#ab7349; RRID:AB_305869 |
O1 | CCF Hybridoma Core Facility | N/A |
O4 | CCF Hybridoma Core Facility | N/A |
ASCL2 | Millipore | Cat#MAB4417; RRID:AB_10562654 |
GFAP | Agilent/Dako | Cat#Z033401-2 |
Donkey anti-rat AF488 | Thermo Fisher Scientific | Cat#A-21208; RRID:AB_2535794 |
Donkey anti-mouse AF488 | Thermo Fisher Scientific | Cat# A-21202; RRID:AB_141607 |
Donkey anti-mouse AF647 | Thermo Fisher Scientific | Cat#A31571; RRID:AB_162542 |
Donkey anti-rabbit AF647 | Thermo Fisher Scientific | Cat#A31573; RRID:AB_2536183 |
HIF1a | Abcam | Cat#Ab2185; RRID:AB_302883 |
Phospho-p44/42. (phosphorylated-ERK1/2) | Cell Signaling Technologies | Cat#9101S; RRID:AB_331646 |
P44/42 (total ERK1/2) | Cell Signaling Technologies | Cat#9102S; RRID:AB_330744 |
Phospho-ERK5 (Thr218/Tyr220) | Cell Signaling Technologies | Cat#3371S; RRID:AB_2140424 |
ERK5 (total) | Cell Signaling Technologies | Cat#3372S; RRID:AB_330491 |
DLX3 | Abcam | Cat#Ab178428 |
SOX10 | R&D Systems | Cat#AF2864; RRID:AB_442208 |
B-Actin peroxidase | Sigma Aldrich | Cat#A3854; RRID:AB_262011 |
VHL | BD Biosciences | Cat#556347; RRID:AB_396376 |
MBP | Abcam | Cat#ab7349; RRID:AB_305869 |
MBP | BioLegend | Cat#808401; RRID:AB_2564741 |
MAG | Thermo Fisher Scientific | Cat#346200; RRID:AB_2533179 |
H3K27Ac | Abcam | Cat#Ab4729; RRID:AB_2118291 |
MyRF | Gift from Michael Wegner | |
HA | Cell Signaling Technologies | Cat#3724S; RRID:AB_1549585 |
Normal Rabbit IgG | Cell Signaling Technologies | Cat#2729S; RRID:AB_1031062 |
SOX6 | abcam | Cat#30455; RRID:AB_1143033 |
A2B5 | Millipore | Cat#MAB312; RRID:AB_94709 |
NKX2-2 | DSHB | Cat#74.5A5; RRID:AB_531794 |
Bacterial and Virus Strains | ||
Stbl3 E.Coli | ThermoFisher | Cat#C737303 |
Lenti-X shots for lenti-virus generation | Clonetech | Cat# 631276 |
Biological Samples | ||
MCH forebrain mouse protein lysate samples | Gift from Brian Popko | N/A |
Chemicals, Peptides, and Recombinant Proteins | ||
N2 Max | R&D Systems | Cat#AR009 |
B27 | Thermo Fisher Scientific | Cat#12587010 |
bFGF | R&D Systems | Cat#23-3FB-01M |
PDGFA | Thermo Fisher Scientific | Cat#221-AA-050 |
DMEM/F12 | Thermo Fisher | Cat#11320-033 |
DMEM | Thermo Fisher | Cat# 11960-044 |
Fetal Bovine Serum (FBS) | Fisher | Cat#A3160402 |
MEM Non-essential amino acids | Thermo Fisher | Cat#11140-050 |
Glutamax | Thermo Fisher | Cat#35050061 |
2-Mercaptoethanol | Sigma | Cat#M3148 |
Poly-L-ornithine | Sigma Aldrich | Cat#P3655-50MG |
Laminin | Sigma Aldrich | Cat#L2020-1MG |
Noggin | R&D Systems | Cat#3344NG050 |
IGF-1 | R&D Systems | Cat#291G1200 |
cyclic AMP | Sigma Aldrich | Cat#D0260-100MG |
NT3 | Thermo Fisher Scientific | Cat#267N3025 |
T3 (Thyroid hormone) | Sigma Aldrich | Cat#T-6397 |
Hb-EGF | R&D Systems | Cat#259-HE-050 |
CNTF | R&D Systems | Cat#557-NT-010 |
BMP4 | R&D Systems | Cat#314-BP-050 |
CloneR | Stem Cell Technologies | Cat#5889 |
LDN193189 | Sigma | Cat#SML0559 |
SB-43152 | Sigma | Cat#S4317 |
Normal donkey serum | Jackson ImmunoResearch | Cat#017-000-121 |
Paraformaldehyde | Electron microscopy sciences | Cat#15710 |
Triton X-100 | Sigma Aldrich | Cat#1086431000 |
DAPI | Sigma Aldrich | Cat#D8417 |
Puromycin | Thermo Fisher Scientific | Cat#A1113802 |
Blasticidin | Thermo Fisher Scientific | Cat#A1113903 |
Hygromycin | Thermo Fisher Scientific | Cat#10687010 |
SCH772984 | Selleck Chemicals | Cat#S7101 |
AZD0364 | Selleck Chemicals | Cat#S8708 |
VX-11e | Selleck Chemicals | Cat#S7709 |
BIX02188 | Selleck Chemicals | Cat#S1530 |
BIX02189 | Selleck Chemicals | Cat#S1531 |
RIPA Buffer | Sigma | Cat# R0278 |
Protease and Phosphatase Inhibitor | Thermo Fisher Scientific | Cat#78441 |
NuPAGE Bis-Tris gels | Thermo Fisher Scientific | Cat#NP0335BOX |
PVDF membranes | Thermo Fisher Scientific | Cat#LC2002 |
TRIzol | Ambion | Cat#15596-026 |
Protein G magnetic DynaBeads | Thermo Fisher Scientific | Cat#10004D |
Proteinase K | Thermo Fisher | Cat#AM2546 |
Ampure XP beads | Aline Biosciences | Cat#C-1003-5 |
Hypoxyprobe-1 | Hypoxyprobe Inc | Cat#HP1-100Kit |
Penicillin-streptomycin | ThermoFisher | Cat#15070063 |
Bioactive Compound Library-I | Selleck Chemicals | Cat#1700 |
Critical Commercial Assays | ||
QIAquick Gel Extraction Kit | Qiagen | Cat#28115 |
Qiagen Plasmid Maxi Kit (25) | Qiagen | Cat#12163 |
KAPA HiFi HotStart ReadyMix | KAPA Biosystems | Cat#kk2602 |
KAPA Sybr Fast qPCR kit | KAPA Biosystems | Cat# KK4605 |
Taqman gene expression master mix | Thermo Fisher | Cat#4369016 |
NEBNext® Library Quant Kit for Illumina® | New England 641 Biolabs | Cat#E7630 |
Protein Assay Kit | Bio-Rad | Cat#5000001 |
RNeasy Mini Kit | Qiagen | Cat#74104 |
iScript cDNA synthesis kit | Bio-Rad | Cat#1708891 |
NEBNext Poly(A) mRNA Magnetic Isolation Module | NEB | Cat#E7490L |
NEBNext Ultra RNA Library Prep Kit for Illumina | NEB | Cat#E7530L |
truChIP Chromatin Shearing Kit with Formaldehyde | Covaris | Cat#520154 |
Hypoxyprobe Kit | Hypoxyprobe Inc | Cat#HP1-100Kit |
ON-TARGETplus Mouse siRNA smartpool: Ascl2 | Horizon/Dharmacon | Cat#L-047262-01-0005 |
ON-TARGETplus Mouse siRNA smartpool: Dlx3 | Horizon/Dharmacon | Cat#L-041957-01-0005 |
siGENOME Mouse siRNA pool: Non-Targeting Control #2 | Horizon/Dharmacon | Cat#D-001206-14-05 5 nmol |
Basic NucleofectorTM Kit for Primary Mammalian Glial Cells | Lonza | Cat# VPI-1006 |
Qiagen MinElute PCR purification Kit | Qiagen | Cat#28004 |
MTS assay kit | abcam | Cat# ab197010 |
Deposited Data | ||
RNA-seq datasets | This paper | GSE143474; GSE143473 (subseries) |
ChIP-seq datasets | This paper | GSE143474; GSE143472 (subseries) |
ATAC-seq datasets | This paper | GSE143474; GSE155366 (subseries) |
Experimental Models: Cell Lines | ||
Lenti-X 293T Cell line | Takara Biosciences | Cat#632180 |
Epiblast stem cell (EpiSC) derived OPCs | Najm et al. 2011 | N/A |
Human iPSCs (H7) to derive Oligocortical spheroids | NIH hESC registry | NIHhESC-10-0061; RRID:CV-CL_9772 |
Experimental Models: Organisms/Strains | ||
Mouse, Male: 129S/SvEv E3.5 for EpiSC isolation | Jackson Laboratory | RRID:MGI:3050593 |
Mouse, mixed sex, postnatal day 2 and 7: C57BL/6J | Jackson Laboratory | RRID:IMSR_JAX:000664 |
Oligonucleotides | ||
Primer: sgVHL Forward: TCCCTACACGACGCTCTTCCGATCTCTCTCAGGTCATCTTCTGCAACC | This manuscript | IDT |
Primer: sgVHL Reverse: AGTTCAGACGTGTGCTCTTCCGATCTGACAAGATGCTCGGGGTCGG | This manuscript | IDT |
Primer: sgVHL.2 Forward: TCCCTACACGACGCTCTTCCGATCTAATAAACAGGTGCCATGCCC | This manuscript | IDT |
Primer: sgVHL.2 Reverse: AGTTCAGACGTGTGCTCTTCCGATCTAGATTGACTATTAACCTGGCAATG | This manuscript | IDT |
sgRNAs | See methods | IDT |
Taqman probes | See methods | IDT |
Recombinant DNA | ||
CRISPRv2 constructs | Sanjana et al., 2014 | Addgene plasmid #52961; RRID:Addgene_52961 |
lentiSAMv2 constructs | Joung et al., 2017 | Addgene plasmid #75112; RRID:Addgene_75112 |
Activation helper | Joung et al., 2017 | Addgene plasmid #89308; RRID:Addgene_89308 |
N-terminus tagged 3x HA Ascl2 construct | Genecopoeia | Cat# EX-Mm03774-Lv117 |
Software and Algorithms | ||
Columbus Image Data Storage and Analysis System | Perkin Elmer | www.perkinelmer.com/Product/columbus |
Outknocker | Schmid-Burgk et al., 2014 | http://www.outknocker.org/outknocker2.htm |
Graphpad Prism | Graphpad Software | https://www.graphpad.com/scientific-software/prism/; RRID: SCR_002798 |
Image Studio Acquisition software | LI-COR | LI-COR Image Studio Software; RRID:SCR_015795 |
Salmon 0.14.1 | Patro et al. 2017 | https://github.com/COMBINE-lab/salmon |
DESEQ2 | Love et al. 2014 | https://bioconductor.org/packages/release/bioc/html/DESeq2.html; RRID:SCR_015687 |
Metascape | Zhou et al. 2019 | http://metascape.org/gp/index.html#/main/step1; RRID:SCR_016620 |
GSEA | Subramanian et al. 2005 | https://www.gsea-msigdb.org/gsea/index.jsp; RRID:SCR_003199 |
Trim Galore! Version 0.4.1 | Krueger, F. 2015 | http://www.bioinformatics.babraham.ac.uk/projects/trim_galore/; RRID:SCR_011847 |
Bowtie2 Version 2.3.2 | Langmead and Slazberg, 2012 | http://bowtie-bio.sourceforge.net/bowtie2/index.shtml; RRID:SCR_005476 |
MACs Version 2.1.1 | Feng et al. 2012 | http://liulab.dfci.harvard.edu/MACS/; RRID:SCR_013291 |
Integrative Genomics Viewer | Broad Institute | http://www.broadinstitute.org/igv/; RRID:SCR_011793 |
Galaxy | Afgan et al. 2018 | http://galaxyproject.org/; RRID:SCR_006281 |
Diffbind software | Ross-Innes et al. 2012 | https://bioconductor.org/packages/release/bioc/html/DiffBind.html; RRID:SCR_012918 |
HOMERv4.11.1 | Heinz et al., 2010 | http://homer.ucsd.edu/; RRID:SCR_010881 |
BEDTools Version 2.25.0 | Quinlan and Hall, 2010 | https://github.com/arq5x/bedtools2; RRID:SCR_006646 |
Deeptools Version 3.3.1 | Ramirez et al., 2016 | https://deeptools.readthedocs.io/en/develop/; RRID:SCR_016366 |
Microsoft Excel | Microsoft | 2016 |
Illustrator CC | Adobe | 2020 |
Other | ||
C-Chamber | BioSpherix | Cat#C-274 |
96-well plate | ThermoFisher | Cat#167008 |
384-well PDL-coated plate | PerkinElmer | Cat#6057500 |
Operetta High Content Imaging and Analysis System | PerkinElmer | N/A |
EL406 Microplate Washer Dispenser | BioTek | N/A |
5μl dispense cassette | BioTek | N/A |
Janus automated workstation | PerkinElmer | N/A |
Odyssey Fc Imaging System | LI-COR | N/A |
7300 real-time PCR system | Applied Biosystems | N/A |
5973 Mass Spectrometer | Agilent | N/A |
Hamamatsu Nanozoomer S60 Slide Scanner | Hamamatsu | C13210-01 |
SuperSignal West Pico PLUS Chemiluminescent Substrate | ThermoFisher | Cat#34580 |
Halt Protease and Phosphatase Inhibitor Cocktail, EDTA-free (100x) | ThermoFisher | Cat#78441 |
Nextera Tn5 Transposase | Illumina | Cat#FC-121-1030 |
Covaris S2 Focused Ultrasonicator | Covaris | Cat#17015 |
Amaxa Nucleofector 2b | Lonza | Cat#AAB-1001 |
E12.5 heart HIF1a ChIP-Seq | Guimaraes-Camboa et al., 2015 | GSM1500750, GSM1500751, GSM1500749 |
Melanocyte HIF1a ChIP-Seq | Loftus et al., 2017 | GSM2305570, GSM2305571, GSM2305572 |
Th17 T-cell HIF1a ChIP-Seq | Ciofani et al., 2012 | GSM1004819, GSM1004991, GSM1004820, GSM1004993 |
Mus musculus mm10 genome build | NCBI; Mouse Genome Sequencing Consotium | https://www.ncbi.nlm.nih.gov/assembly/GCF_000001635.20/ |