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 |