Antibodies |
anti-ATRX |
Santa Cruz Biotechnology |
sc-15408 |
anti-ATR |
Santa Cruz Biotechnology |
sc-51573 |
anti-ATRX |
Bethyl Laboratories, LLC |
A301-045A |
anti-Vinculin |
Invitrogen |
#700062 |
anti-CHK1 |
Cell Signaling Technology |
#2360 |
anti-pCHK1 Ser345 |
Cell Signaling Technology |
#2348 |
anti-CHK2 pT68 |
Cell Signaling Technology |
#2661 |
anti-Phospho-Histone H2A.X Ser139 |
Cell Signaling Technology |
#2577 |
anti-mouse IgG, HRP-linked Antibody |
Cell Signaling Technology |
#7076 |
anti-ATM pS1981 |
Abcam |
ab81292 |
HRP Goat Anti-Rabbit IgG Antibody |
Vector Laboratories |
P1-1000 |
Anti-Histone H3.3 Antibody |
Millipore Sigma |
#09838 |
Control IgG |
Millipore Sigma |
#12370 |
Alexa-Fluor 488 |
ThermoFisher |
A-11034 |
Rabbit Phospho-Histone H3 (Ser10) primary
antibody |
Cell Signaling |
3377T |
Goat Anti-Rabbit IgG (H+L) FITC-conjugated
secondary antibody |
ThermoFisher |
F2765 |
Chemicals, Peptides, and
Recombinant Proteins |
B-27 supplement |
Gibco |
12587-010 |
N-2 supplement |
Gibco |
17502-048 |
Penicillin-Streptomycin |
Cellgro |
30-001-CI |
Normocin |
Invivogen |
ant-nr-1 |
FGF |
Shenandoah Biotech |
100-26 |
EGF |
Shenandoah Biotech |
100-146 |
GlutaMAX-I Supplement |
Invitrogen |
#35050061 |
Fetal Bovine Serum (FBS) |
Sigma |
F4135-500ML |
Clarity™ Western ECL substrate |
Bio-rad |
#170-5060 |
DMSO |
Sigma |
D8418-250ML |
Paraformaldehyde (PFA) |
Sigma |
158127-500G |
Prolong™ Gold antifade reagent with
DAPI |
Invitrogen |
P36931 |
AZD0156 |
Selleck |
S8375 |
AZD1390 |
Selleck |
S8680 |
KU-55933 |
Selleck |
S1092 |
KU-60019 |
Selleck |
S1570 |
Nocodazole |
Sigma |
M1404-10MG |
Triton X-100 |
Sigma |
T8787 |
Propidium Iodide |
Sigma |
P4864 |
RNase A |
Qiagen |
19101 |
Formic acid |
Fisher Scientific |
60-006-16 |
D-luciferin |
Gold Biotechnology |
#115144-35-9 |
Critical Commercial Assays |
Qiagen RNeasy Mini Kit |
Qiagen |
#74104 |
TruSeq Stranded mRNA Library Kit |
Illumina |
#20020594 |
DNeasy Blood and Tissue Kit |
Qiagen |
#69504 |
Deposited Data |
Raw data |
this paper |
GSE178116 |
Atrx inactivation drives disease-defining
phenotypes in glioma cells of origin through global epigenomic
remodeling |
Danussi et
al., 2018
|
GSE100462, GSE100464
|
Integrated Molecular Meta-Analysis of 1,000
Pediatric High-Grade and Diffuse Intrinsic Pontine Glioma |
Mackay et al.,
2017
|
EGA - EGAS00001000226, EGAS0000100192,
EGAS00001000575, EGAS00001000720, EGAS00001001139; the Gene Expression
Omnibus (www.ncbi.nlm.nih.gov/geo/)-GSE19578, GSE26576, GSE21420, GSE34824, GSE36245, GSE36278
GSE50022, GSE50021, GSE50024, GSE55712; ArrayExpress - E-TABM-857, E-TABM-1107. |
Genomic Data Commons (GDC) Data Portal |
The Cancer Genome Atlas Program |
https://portal.gdc.cancer.gov
|
single cell RNA-seq data of oligodendroglioma
samples |
Tirosh et al.,
2016b
|
GSE7063 |
single cell RNA-seq data of astrocytomas
samples |
Venteicher et
al., 2017
|
GSE89567 |
Experimental Models: Cell
Lines |
KNS42 |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
SJ-GBM2 |
Children’s Oncology Group cell line
repository |
#12440053 |
U251 ATRXEV |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
U251 ATRXKO |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
SF188 ATRXEV |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
SF188 ATRXKO |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
UW479 ATRXEV |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
UW479 ATRXKO |
Dr. Alan Meeker, Johns Hopkins University,
Baltimore, MD |
N/A |
Human GBM D54 ATM-Luciferase inhibition
reporter cells |
Dr. Alnawaz Rehemtulla University of Michigan,
Ann Arbor, MI |
N/A |
U251 ATRXKO Chk1EV |
This paper |
N/A |
U251 ATRXKO Chk1OE |
This paper |
N/A |
U251 ATRXKO FastFUCCI |
This paper |
N/A |
U251 ATRXEV FastFUCCI |
This paper |
N/A |
U251 ATRXKO Chk1EV FastFUCCI |
This paper |
N/A |
U251 ATRXKO Chk1OE FastFUCCI |
This paper |
N/A |
UW479 ATRXKO Chk1EV |
This paper |
N/A |
UW479 ATRXKO Chk1KO |
This paper |
N/A |
SF188 ATRXKO Chk1EV |
This paper |
N/A |
SF188 ATRXKO Chk1KO |
This paper |
N/A |
Experimental Models:
Organisms/Strains |
Mouse: C57BL/6 |
Charles River or Jackson Labs |
N/A |
Mouse: NOD scid gamma (NSG) |
Charles River or Jackson Labs |
N/A |
Mouse: FVB |
Charles River or Jackson Labs |
N/A |
Oligonucleotides |
Human Chk1 Forward Primer
5′GGTGCCTATGGAGAAGTTCAA |
IDT |
N/A |
Human Chk1 Reverse Primer
5′TCTACGGCACGCTTCATATC |
IDT |
N/A |
Human GAPDH Forward Primer
5′GGATTTGGTCGTATTGGG |
IDT |
N/A |
Human GAPDH Reverse Primer
5′GGAAGATGGTGATGGGATT |
IDT |
N/A |
Primers (see Table S7) |
This paper |
N/A |
Recombinant DNA |
shp53 lentiviral plasmid |
Dr. John Ohlfest, University of Minnesota,
Minneapolis, Minnesota |
N/A |
shATRX lentiviral plasmid |
generated for Koschmann et al., 2016 |
N/A |
NRAS overexpression plasmid |
Dr. John Ohlfest, University of Minnesota,
Minneapolis, Minnesota |
N/A |
pcDNA4-Chk1-Flag Overexpression Plasmid |
Addgene |
#22894 |
pBOB-EF1-FastFUCCI-Puro Plasmid |
Addgene |
#86849 |
Software and Algorithms |
chipenrich (v2.6.1) |
Welch et al.,
2014
|
https://bioconductor.org/packages/release/bioc/html/chipenrich.html
|
annotatr (v1.0.3) |
Cavalcante and
Sartor, 2017
|
https://www.bioconductor.org/packages/release/bioc/html/annotatr.html
|
Bowtie2 (v2.2.1) |
Langmead and
Salzberg, 2012
|
http://bowtie-bio.sourceforge.net/bowtie2/index.shtml
|
deepTools (v.3.1.3) |
Ramirez et
al., 2016
|
https://deeptools.readthedocs.io/en/develop/
|
IGV (v2.11.3) |
Robinson et
al., 2011
|
https://igv.org
|
FastQC (v0.11.5) |
Andrews, S., 2010 |
https://www.bioinformatics.babraham.ac.uk/projects/fastqc/
|
TrimGalore (v0.6.0) |
Github |
https://github.com/FelixKrueger/TrimGalore
|
STAR (v2.5.3a) |
Dobin et al.,
2013
|
https://github.com/alexdobin/STAR
|
HTSeq (v0.11.2) |
Anders et al., 2015
|
https://htseq.readthedocs.io/en/master/
|
edgeR (v3.20.9) |
Robinson et
al., 2010
|
https://bioconductor.org/packages/release/bioc/html/edgeR.html
|
RNA-Enrich |
N/A |
http://lrpath.ncibi.org
|
betareg
|
Grün et al., 2011
|
https://cran.r-project.org/web/packages/betareg/betareg.pdf
|
Seurat (v4.0.5) |
Stuart et al.,
2019
|
https://satijalab.org/seurat/
|
fgsea |
Korotkevich et
al., 2019
|
https://bioconductor.org/packages/release/bioc/html/fgsea.html
|