REAGENT or RESOURCE | SOURCE | IDENTIFIER |
Antibodies | ||
HA (WB) | Biolegend | Cat# 901501 ; RRID: AB_2565006 |
H3K27M (WB and immunohistochemistry) | EMD | Cat# ABE419 ; RRID: AB_2728728 |
H3K27me3 (WB) | EMD | Cat# 07–449 ; RRID: AB_310624 |
H3K27ac (WB) | abcam | Cat# ab177178 ; RRID: AB_2828007 |
H3K4me1 (WB) | Cell signaling | Cat# 5326s ; RRID: AB_10695148 |
H3K4me3 (WB) | Cell signaling | Cat# 9751s ; RRID: AB_2616028 |
H3K36me3 (WB) | ACTIVE MOTIF | Cat# 61021 ; RRID: AB_2614986 |
Total H3 (WB) | Cell signaling | Cat# 3638S ; RRID: AB_1642229 |
H3K27me3 (ChIP-seq) | ACTIVE MOTIF | Cat# 39155 ; RRID: AB_2561020 |
H3K27ac (ChIP-seq) | ACTIVE MOTIF | Cat# 39133 ; RRID: AB_2561016 |
H3K4me1 (ChIP-seq) | ACTIVE MOTIF | Cat# 39297 ; RRID: AB_2615075 |
H3K4me3 (ChIP-seq) | ACTIVE MOTIF | Cat# 39159 ; RRID: AB_2615077 |
H3K27ac (immunohistochemistry) | Cell signaling | Cat# 8173 ; RRID: AB_10949503 |
H3K27M (immunohistochemistry) | EMD | Cat# ABE419 ; RRID: AB_2728728 |
H3K27me3 (immunohistochemistry) | EMD | Cat# 07–449 ; RRID: AB_310624 |
SLC1A5 (immunohistochemistry) | Sigma | Cat# HPA035240 ; RRID: AB_10604092 |
SLC2A3/GLUT3 (WB) | abcam | Cat# ab191071 ; RRID: AB_2736916 |
HK2 (WB) | Santa Cruz bio | Cat# sc-374091;RRID:AB_10917915 |
IDH1 (WB) | Cell signaling | Cat# 8137 ; RRID: AB_10950504 |
IDH2 (WB) | Santa Cruz bio | Cat# sc-374476; RRID: AB_10986415 |
GDH (GLUD1/2) (WB) | abcam | Cat# ab166618 ; RRID: |
SLC7A11 (WB) | Cell signaling | Cat# 12691s ; RRID: AB_2687474 |
β-ACTIN (WB) | Sigma | Cat# A5441 ; RRID: AB_476744 |
GAPDH (WB) | Santa Cruz bio | Cat# sc-25778; RRID: AB_10167668 |
p-s6rp (Ser240/244) (WB) | Cell signaling | Cat# 2215s ; RRID: AB_331682 |
p-s6rp (Ser235/236) (WB) | Cell signaling | Cat# 4858s ; RRID: AB_916156 |
s6rp (WB) | Cell signaling | Cat# 2317s ; RRID: AB_2238583 |
N-myc (WB) | Santa Cruz bio | Cat# sc-53993; RRID: AB_831602 |
C-myc (WB) | Santa Cruz bio | Cat# sc-40 ; RRID: AB_627268 |
SLC1A5/ASCT2 (WB) | ThermoFisher | Cat# PA5–50527;RRID: AB_2635980 |
GLS (WB) | abcam | Cat# ab156876 ; RRID: AB_2721038 |
GSK3 a/b (WB) | Cell signaling | Cat# 5676s ; RRID: AB_10547140 |
p-GSK3 a/b (Ser21/9) (WB) | Cell signaling | Cat# 9331s ; RRID: AB_329830 |
VINCULIN (WB) | Sigma | Cat# V9264 ; RRID: AB_10603627 |
FLAG (WB) | Cell signaling | Cat# 2368s ; RRID: AB_2217020 |
GOT1 (WB) | Abcam | Cat# ab171939 |
Hif-1α (WB) for mouse | Abcam | Cat# ab16066; RRID:AB_302234 |
HIF-1α (WB) for human | Abcam | Cat# ab179483; RRID:AB_2732807 |
Bacterial and Virus Strains | ||
One-shot Stbl3 chemically competent E.coli | ThermoFisher | Cat# C737303 |
Chemicals, Peptides, and Recombinant Proteins | ||
DM-succinate | Acros Organics | Cat# AC150731000 |
DM-αKG | Sigma | Cat# 349631 |
DMSO | Sigma | Cat# D2650 |
Glucose | Gibco | Cat# A2494001 |
L-Glutamine | Gibco | Cat# 25030–081 |
Matrix-gel (Matrigel® Basement Membrane Matrix) | Corning | Cat# 356234 |
D-Luciferin, Potassium Salt | GoldBio | Cat# LUCK-1G |
GSK-J4 | Cayman Chem. | Cat# 12073 |
GSK-J5 | Cayman Chem. | Cat# 12074 |
DON (6-Diazo-5-oxo-L-norleucine) | Cayman Chem. | Cat# 17580 |
2-DG (2-Deoxy-D-glucose) | Cayman Chem. | Cat# 14325 |
D-2HG ((2R)-Octyl-α-hydroxyglutarate) | Cayman Chem. | Cat# 16366 |
CB-839 | Cayman Chem. | Cat# 22038 |
BPTES | Cayman Chem. | Cat# 19284 |
Lonidamine | Cayman Chem. | Cat# 14640 |
Ketoconazole | Cayman Chem. | Cat# 15212 |
Posaconazole | Cayman Chem. | Cat# 14737 |
Desferrioxamine | Cayman Chem. | Cat# 14595 |
JHU-083 | MCE | Cat# HY-122218 |
IDH1 inhibitor 1 | Abbvie | |
IDH1 inhibitor 2 | Abbvie | |
IDH1 inhibitor 3 | Abbvie | |
IDH1 inhibitor 13 | Abbvie | |
Inactive enone analogue 18 | Abbvie | |
Critical Commercial Assays | ||
TransIT®-Lenti Transfection Reagent | Mirus | Cat# MIR 6650 |
TMT10plex™ Isobaric Label Reagent Set | ThermoFisher | Cat# 90110 |
D-2-Hydroxyglutarate Assay Kit | abcam | Cat# ab211070 |
Succinate Assay Kit (Colorimetric) | abcam | Cat# ab204718 |
Alpha Ketoglutarate (alpha KG) Assay Kit | abcam | Cat# ab83431 |
Deposited Data | ||
NCBI/ GEO (GSE135419), next generation sequencing files. | This paper | GEO: GSE135419 |
Experimental Models: Cell Lines | ||
HSJD-DIPG007 (H3.3K27M) | Dr. Rintaro Hashizume, Northwestern University; RRID: CVCL_VU70 | |
SU-DIPG-XIII (H3.3K27M) | Dr. Michelle Monje, Stanford University; RRID: CVCL_IT41 | |
SU-DIPG-IV (H3.1K27M) | Dr. Michelle Monje, Stanford University; RRID: CVCL_IT39 | |
SF7761 (H3.3K27M) | Dr. Rintaro Hashizume, Northwestern University; RRID: CVCL_IT45 | |
SJGBM2 (H3WT) | Dr. Carl Koschmann, University of Michigan; RRID: CVCL_M141 | |
SF188 (H3WT) | Dr. Craig B. Thompson, Memorial Sloan Kettering Cancer Center; RRID: CVCL_6948 | |
UMPed37 (H3WT) | Dr. Carl Koschmann, University of Michigan | |
KNS42 (H3.3G34V) | Dr. Carl Koschmann, University of Michigan; RRID: CVCL_0378 | |
TB-096 (IDH1 R132H) | Dr. Hai Yan, Duke University | |
mNSC | Dr. Richard J. Gilbertson, St Jude’s Hospital | |
Experimental Models: Organisms/Strains | ||
Mouse: Hsd:Athymic Nude-Foxn1nu | Jackson Laboratories | Cat# 002019; RRID: IMSR_JAX:002019 |
Mouse: NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ | Jackson Laboratories | Cat#:005557; RRID: IMSR_JAX:005557 |
Oligonucleotides | ||
siRNA IDH1 NT: ON-TARGETplus Non-targeting Control siRNAs | Dharmacon | Cat# D-001810–01-05 |
siRNA IDH1–1: ON-TARGETplus IDH1 siRNA Targeted Region:3'UTR | Dharmacon | Cat# J-008294–09-0002 |
siRNA IDH1–2: ON-TARGETplus IDH1 siRNA Targeted Region:ORF | Dharmacon | Cat# J-008294–10-0002 |
siRNA IDH1–3: ON-TARGETplus IDH1 siRNA Targeted Region:ORF | Dharmacon | Cat# J-008294–11-0002 |
Recombinant DNA | ||
H3.3WT lentiviral plasmid | Dr. David Allis, The Rockefeller University | |
H3.3K27M lentiviral plasmid | Dr. David Allis, The Rockefeller University | |
shNT lentiviral plasmid | Sigma | Cat# SHC002V |
shGLUD1–1 lentiviral plasmid | Sigma | Cat# TRCN0000343656 |
shGLUD1–2 lentiviral plasmid | Sigma | Cat# TRCN0000028600 |
shHK2–1 lentiviral plasmid | Sigma | Cat# TRCN0000037670 |
shHK2–2 lentiviral plasmid | Sigma | Cat# TRCN0000195582 |
shIDH1 lentiviral plasmid (V2LHS_217815) | Dharmacon | Cat# RHS4430–200189861 |
pLenti PGK Blast V5-LUC | Addgene | Cat# 19166 ; RRID: Addgene_19166 |
pSLIK-IDH1-FLAG | Addgene | Cat# 66802 ; RRID: Addgene_66802 |
pSLIK-IDH1-R132H-FLAG | Addgene | Cat# 66803 ; RRID: Addgene_66803 |
Software and Algorithms | ||
PRISM 7.0/8.0 | GraphPad | |
Image Studio software | LI-COR | |
IVIS Living Image Software | Perkin Elmer | |
IGV(Integrative Genomics Viewer) 2.5.0 | Broad Institute | |
AperioImageScope/ AperioScan Scope | Aperio | |
RSEM (RNA-Seq by Expectation-Maximization) | https://deweylab.github.io/RSEM/ | |
CASAVA 1.8.2 pipeline | Illumina | |
methylKit | https://bioconductor.org/packages/release/bioc/html/methylKit.html | |
Proteome Discoverer v2.1 | Thermo Fisher | |
SICER | http://www.lcmodel.com/ | |
MACS | http://liulab.dfci.harvard.edu/MACS/ | |
Agilent MassHunter Workstation Software Quantitative | Agilent | |
GSEA | Broad Institute http://software.broadinstitute.org/gsea/index.jsp |
|
MetaboAnalyst | https://www.metaboanalyst.ca/ | |
Matlab’s image processing toolbox | Matlab | |
Image Studio software | Licor |