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. 2022 Oct 10;40(10):1111–1127.e9. doi: 10.1016/j.ccell.2022.08.014
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies

InVivoMAb anti-mouse CD8α (clone 2.43) BioXCell Cat #: BE0061; RRID: AB_1125541
InVivoMAb anti-mouse CD4 (clone GK1.5) BioXCell Cat #: BE0003-1; RRID: AB_1107636
InVivoPlus anti-mouse IFNγ (clone XMG1.2) BioXCell Cat #: BP0055; RRID: AB_1107694
InVivoMAb anti-mouse VEGFR-2 (clone DC101) BioXCell Cat #: BE0060; RRID: AB_1107766
InVivoMAb anti-mouse CXCR3 (CD183) (clone CXCR3-173) BioXCell Cat #: BE0249; RRID: AB_2687730
InVivoMAb anti-mouse/human VLA-4 (CD49d) (clone PS/2) BioXCell Cat #: BE0071; RRID: AB_1107657
InVivoMAb anti-mouse PD-L1 (B7-H1) BioXCell Cat #: BE0101; RRID: AB_10949073
FITC TNF alpha Rat anti-mouse (cloneMP6-XT22) eBioscience Cat #: 11-7321-82; RRID: AB_465418
PE Granzyme B anti-mouse (clone NGZB) eBioscience Cat #: 12-8898-82; RRID: AB_10870787
PerCP-Cyanine5.5 CD3e anti-mouse (clone 145-2C11) eBioscience Cat #: 45-00310-82; RRID: AB_1107000
PE/Cyanine7 CD8a anti-mouse (clone 53-6.7) Biolegend Cat #: 100722; RRID: AB_312760
Biotin CD11b anti-mouse (clone M1/70) eBioscience Cat#: 13-0112-82; RRID: AB_466359
APC IFN-γ anti-mouse (clone XMG1.2) Biolegend Cat #: 505810; RRID: AB_315403
APC/Cyanine7 CD45 anti-mouse (clone 30-F11) Biolegend Cat#: 103116; RRID: AB_312980
FITC CD62L anti-mouse (clone MEL-14) Biolegend Cat#: 104406; RRID: AB_313093
PerCP-Cyanine5.5 CD11b anti-mouse (clone M1/70) eBioscience Cat#: 45-0112-82; RRID: AB_953558
Pacific Orange™ CD8a anti-mouse (clone 5H10) Invitrogen Cat#: MCD0830, RRID: AB_10376311
APC CD45 anti-mouse (clone 30-F11) BD Biosciences Cat#: 559864; RRID: AB_398672
Pe-Cyanine7 CD3e (clone 145-2C11) eBioscience Cat#: 25-0031-81; RRID: AB_469572
Alexa Fluor® 488 Anti-Stat5 (pY694) anti-mouse (clone 47/Stat5(pY694)) BD Biosciences Cat#: 612598; RRID: AB_399881
PE TCF-7/TCF-1 anti-mouse (clone S33-966) BD Biosciences Cat#: 564217; RRID: AB_2687845
Pacific Blue™ Ki-67 anti-mouse Biolegend Cat#: 652422; RRID: AB_2564490
PerCP-Cyanine5.5 Foxp3 anti-mouse (clone FJK-16s) eBioscience Cat#: 45-5773-82; RRID: AB_914351
APC CD3e anti-mouse (clone 145-2C11) Biolegend Cat#: 100312; RRID: AB_312677
Biotin CD4 anti-mouse (clone GK1.5) Biolegend Cat#: 100404; RRID: AB_312689
Pe-Cyanine7 CD4 anti-mouse (clone RM4-5) Biolegend Cat#: 100528; RRID: AB_312729
Brilliant Violet 510™ CD11b anti-mouse/human (clone M1/70) Biolegend Cat#: 101245; RRID: AB_2561390
Alexa Fluor® 488 CRACC/SLAMF7 anti-mouse R&D Systems Cat#: FAB46281G
Brilliant Violet 421™ LAP (TGF-β1) anti-mouse (clone TW7-16B4) Biolegend Cat#: 141407; RRID: AB_2561580
APC-eFluor™ 780 CD45 anti-mouse (clone 30-F11) Biolegend Cat#: 47-0451-82; RRID: AB_1548781
Brilliant Violet 650™ CD223 (LAG-3) anti-mouse (clone C9B7W) Biolegend Cat#: 125227; RRID: AB_2687209
Brilliant Violet 421™ Eomes anti-mouse (clone X4-83) BD Biosciences Cat#: 567166; RRID: AB_2916483
APC HIF-1 alpha anti-mouse/anti-human (clone 241812) R&D Systems Cat#: IC1935A; RRID: AB_ 1061580
Alexa Fluor™ 488 Arginase 1 anti-mouse/anti-human (clone A1exF5) eBioscience Cat#: 53-3697-82; RRID: AB_2734831
APC/Cyanine7 IL-10 anti-mouse (clone JES5-16E3) Biolegend Cat#: 505036; RRID: AB_2566331
PerCP-Cyanine5.5 Ly-6C anti-mouse (clone HK1.4) eBioscience Cat#: 45-5932-82; RRID: 
AB_2723343
Pacific Blue™ Ly-6G anti-mouse (clone 1A8) Biolegend Cat#: 127612; RRID: AB_2251161
Alexa Fluor® 647 CD49d anti-mouse (clone R1-2) Biolegend Cat#: 103614; RRID: AB_528837
FITC CD49d anti-mouse (clone R1-2) Biolegend Cat#: 103605; RRID: AB_313037
Alexa Fluor® 647 CXCL9 anti-mouse (clone MIG-2F5.5) Biolegend Cat#: 515606; RRID: AB_1877135
FITC I-A/I-E anti-mouse (clone M5/114.15.2) Biolegend Cat#: 107606; RRID: AB_313321
PE CD274 (B7-H1, PD-L1) anti-mouse (clone 10F.9G2) Biolegend Cat#: 124308; RRID: AB_2073556
PE-Cyanine7 CD11b anti-mouse (clone M1/70) eBioscience Cat#: 25-0112-82; RRID: AB_469588
PerCP/Cyanine5.5 CD86 anti-mouse (clone GL-1) Biolegend Cat#: 105028; RRID: AB_2074994
APC IL-6 anti-mouse (clone MP-5-20F3) Biolegend Cat#: 504508; RRID: AB_10694868
PE IL-1α anti-mouse (clone ALF-161) Biolegend Cat#: 503203; RRID: AB_315281
FITC CD8a anti-mouse (clone 53-6.7) Biolegend Cat#: 100706; RRID: AB_312745
Alexa Fluor® 647 CD8a anti-mouse (clone 53-6.7) Biolegend Cat#: 100724; RRID: AB_389326
Alexa Fluor® 647 CD4 anti-mouse (clone RM4-5) Biolegend Cat#: 100530; RRID: AB_389325
FITC CD31 anti-mouse (clone MEC 13.3) BD Biosciences Cat#: 553372; RRID: AB_394818
PE CD31 anti-mouse (clone MEC 13.3) BD Biosciences Cat#: 553373; RRID: AB_394819
Alexa Fluor® 647 CD206 anti-mouse (clone C068C2) Biolegend Cat#: 141712; RRID: AB_10900420
PE F4/80 anti-mouse (clone BM8) eBioscience Cat#: 12-4801-82; RRID: AB_465923
FITC Ki67 anti-mouse (clone 16A8) Biolegend Cat#: 652409; RRID: AB_2562140
MECA-79 Santa Cruz Biotechnology Cat#: sc-19602; RRID: AB_627143
CXCL10/IP-10/CRG-2 anti-mouse R&D Systems Cat#: AF-466-NA; RRID: AB_2292487
Cleaved Caspase-3 (Asp175) (clone 5A1E) Cell Signaling Technology Cat#: 9664; RRID: AB_2070042
CD140b (PDGFRB) anti-mouse (clone APB5) eBioscience Cat#: 14-1402-82; RRID: 
AB_467493
LC3 anti-mouse (clone 5F10) Nanotools Cat#: 0231-100; RRID: AB_2722733
Desmin Abcam Cat#: ab15200; RRID: AB_301744
LAMP1 Abcam Cat#: ab24170; RRID: AB_775978
Arginase I (N-20) Santa Cruz Biotechnology Cat#: sc-18351; RRID: AB_2258542
MMR/CD206 anti-mouse R&D Systems Cat#: AF2535; RRID: AB_2063012
IL-10 anti-mouse (clone  JES052A5) R&D Systems Cat#: MAB417; RRID: AB_2125085
ATG3 Cell Signaling Technology Cat#: 3415; RRID: AB_2059244
HSP-90 (F-8) Santa Cruz Biotechnology Cat#: sc-13119; RRID: AB_675659
GAPDH (clone 14C10) Cell Signaling Technology Cat#: 2118; RRID: AB_561053
Polyclonal Rabbit Anti-Goat Immunoglobulins/HRP antibody DAKO Cat#: P0449; RRID: AB_2617143
Goat Anti-Rabbit Immunoglobulins/HRP antibody DAKO Cat#: P0448; RRID: AB_2617138
Goat Anti-Mouse Immunoglobulins/HRP antibody DAKO Cat#: P0447; RRID: AB_2617137
Anti-rat IgG, HRP-linked antibody Cell Signaling Technology Cat#: 7077; RRID: AB_10694715
Alexa Fluor™ 546 goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody Invitrogen Cat#: A-11035; RRID: AB_2534093
Alexa Fluor™ 647 donkey anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody Invitrogen Cat#: A-21447; RRID: AB_2535864
Alexa Fluor® 647 donkey Anti-Rat IgG H&L Abcam Cat#: ab150155; RRID: AB_2813835
Purified anti-mouse CD16/32 Biolegend Cat#: 101302; RRID: AB_312801
Lycopersicon Esculentum (Tomato) Lectin (LEL, TL), DyLight 594 Invitrogen Cat#: L32471
Lectin Kit I, Biotinylated Vector Laboratories Cat#: BK-1000; RRID: AB_2336252

Bacterial and virus strains

One Shot™ TOP10 Chemically Competent E. coli Invitrogen Cat#: C404010
LVRshp53 second generation lentivirus In house This paper

Chemicals, peptides, and recombinant proteins

Imipramine hydrochloride Sigma-Aldrich Cat#: I0899
(S)-(+)-Clopidogrel hydrogensulfate Sigma-Aldrich Cat#: SML0004
Axitinib (AG 013736) Selleckchem Cat#: S1005
Sunitinib (SU11248) malate Selleckchem Cat#: S1042
Recombinant B20S (anti-VEGF) In house This paper
Desloratadine Sigma-Aldrich Cat#: D1069
Trifluoperazine hydrochloride Sigma-Aldrich Cat#: T6062
HJC0197 Cayman Chemicals Cat#: CAY-19092
CE3F4 Cayman Chemicals Cat#: CAY-17767
Histamine MedChemExpress Cat#: Y-B1204
CFSE Cell Division Tracker Kit Biolegend Cat#: 422701
Dynabeads™ Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation Gibco Cat#: 11452D
Brilliant Violet 421™ Streptavidin Biolegend Cat#: 405226
Streptavidin, Pacific Orange™ conjugate Invitrogen Cat#: S32365
DAPI Roche Cat#: 10236276001
LIVE/DEAD™ Fixable Violet Dead Cell Stain Kit, for 405 nm excitation Invitrogen Cat#: L34964
LIVE/DEAD™ Fixable Red Dead Cell Stain Kit, for 488 nm excitation Invitrogen Cat#: L23102
PrimeScript RT Master Mix Takara Cat#: RR036A
miRNeasy Micro Kit Qiagen Cat#: 217084
miRNeasy Mini Kit Qiagen Cat#: 217004
QuantiNova SYBR Green PCR Kit Qiagen Cat#: 208052
Pierce™ BCA Protein Assay Kit Thermo Scientific™ Cat#: 23225
Fluorescence Mounting Medium DAKO Cat#: S302380
Recombinant Mouse M-CSF (carrier-free) Biolegend Cat#: 576406
ACK Lysing Buffer Gibco Cat#: A1049201
Dispase® II (neutral protease, grade II) Roche Cat#: 04942078001
DNase I recombinant, RNase-free Roche Cat#: 04716728001
Collagenase A Roche Cat#: 10103578001

Critical commercial assays

cAMP Assay Kit (Competitive ELISA) Abcam Cat#: ab65355
EasySep™ Mouse CD11b Positive Selection Kit II StemCell Technologies Cat#: 18970
EasySep™ Mouse CD4+ T Cell Isolation Kit StemCell Technologies Cat#: 19852
EasySep™ Mouse CD8+ T Cell Isolation Kit StemCell Technologies Cat#: 19853
CD31 MicroBeads, mouse Miltenyi Biotec Cat#: 130-097-418
Myelin Removal Beads II, human, mouse, rat Miltenyi Biotec Cat#: 130-096-731
pHrodo™ Green S. aureus Bioparticles™ Conjugate for Phagocytosis Invitrogen Cat#: P35367

Deposited data

TCGA glioblastoma (with AffyU133a array) Goldman et al., 2020 https://xena.ucsc.edu
CHUV EMRs This paper raw data available upon request

Experimental models: Cell lines

293T/17 [HEK 293T/17] ATCC Cat#: CRL-11268™; RRID: CVCL_1926
Mouse GBM-derived cancer cells Derived in house from the LVRshp53 model This paper
DF-1:RCAS-hPDGF-B-HA provided by J.A. Joyce

Experimental models: Organisms/strains

Mouse: FVB.GFAP-Cre In house This paper
Mouse: GFAP-HRasV12; GFAP-CRE; GFAP-LUC; p53flox/wt In house This paper
Mouse: GFAP-HRasV12; GFAP-CRE; GFAP-LUC; p53flox/flox In house This paper
Mouse: C57BL/6J (JAX® Mice Strain) Charles River Strain Code: 632
Mouse: FVB/NCrl Charles River Strain Code: 207
Mouse: Fox Chase SCID Beige Mouse Charles River Strain code: 250
Mouse: iBIP2 (TetO-BRAFv600e, Tyr-CreERT2, Rosa26-Lox-Stop-Lox-rtTA, Pten-fl/fl, Cdkn2a-fl/fl) In house This paper
Mouse: Nestin-Tv-a;Ink4a/Arf−/− (Tg(NES-TVA)J12Ech; Cdkn2atm1Rdp) provided by J.A. Joyce

Oligonucleotides

MISSION® siRNA Universal Negative Control #1 Sigma-Aldrich Cat#: SIC001
MISSION® Predesigned siHrh1 Sigma-Aldrich NM_008285; siRNA ID: SASI_Mm01_00058730
MISSION® Predesigned siHrh1 Sigma-Aldrich NM_008285; siRNA ID: SASI_Mm01_00058731
Primers: see Table S1

Recombinant DNA

ATG3 Mission shRNA plasmid Sigma-Aldrich ID: TRCN0000247440
pMD2G Addgene Cat#: 12259
pCMVR8.74 Addgene Cat#: 22036
pTomo H-rasV12-shp53-Luc provided by I. Verma

Software and algorithms

QuPath (version 0.2.3) Bankhead et al., 2017 https://qupath.github.io
FlowJo (version 10.7.2) BD https://www.flowjo.com
RStudio (version 2022.02.1 Build 461) The R Foundation https://www.r-project.org/
survminer (version 0.4.9) https://cran.r-project.org/web/packages/survminer/index.html
GraphPad Prism (version 9.3.1) Dotmatics https://www.graphpad.com