| Antibodies |
| Anti-mouse PD-1 (clone 29F.1A12, rat, monoclonal) |
Bio X Cell |
Cat# BE0273 |
Immunotherapy treatment |
| RRID:AB_2687796 |
200 μg/mouse twice a week for 2 weeks |
| Anti-mouse G-CSF (clone 67604, rat, monoclonal) |
R&D Systems |
Cat# MAB414 |
Anti G-CSF treatment |
| RRID:AB_2085954 |
10 μg every day for 14 days |
| Anti-GLUT1 (rabbit, polyclonal) |
Millipore |
Cat# 07-1401 |
WB |
| RRID:AB_1587074 |
IF (1:5’000) |
| Anti-GLUT1 (rabbit, monoclonal) |
Abcam |
Cat# ab115730 |
IHC (1:650) |
| RRID:AB_10903230 |
| Anti-Histone H3 (rabbit, polyclonal) |
Abcam |
Cat# ab1791 |
WB |
| RRID:AB_302613 |
| Anti-pHH3 (rabbit, polyclonal) |
Cell Signaling Technology |
Cat# 9701S |
IHC (1:100) |
| RRID:AB_331535 |
| Anti-pErk (rabbit, monoclonal) |
Cell Signaling Technology |
Cat# 4370S |
IHC (1:400) |
| RRID:AB_2315112 |
| Anti-S100A9 (rabbit, polyclonal) |
Novus biologicals |
Cat# NB110-89726 |
IHC (1:5’000) |
| RRID:AB_1217846 |
IF (1:20’000) |
| Anti-CD4 (clone SP35, rabbit, monoclonal) |
Roche Diagnostics |
Cat# 05552737001 |
IF |
| Anti-CD8 (clone SP57, rabbit, monoclonal) |
Roche Diagnostics |
Cat# 05937248001 |
IF |
| Anti-mouse-CD45-BUV661 (clone 30-F11, rat, monoclonal) |
BD Biosciences |
Cat# 565079 |
Flow cytometry |
| RRID:AB_2739057 |
| Anti-mouse/human-CD11b-BV711 (clone M1/70, rat, monoclonal) |
BioLegend |
Cat# 101242 |
Flow cytometry |
| RRID:AB_2563310 |
|
| Anti-mouse-Ly6G-PE (clone 1A8, rat, monoclonal) |
BioLegend |
Cat# 127607 |
Flow cytometry |
| RRID:AB_1186104 |
| Anti-mouse-Siglec-F-PE-Vio 615 (clone REA798, human, monoclonal) |
Miltenyi Biotec |
Cat# 130-112-172 |
Flow cytometry |
| RRID:AB_2653444 |
| Anti-BrdU-APC (clone Bu20a, mouse, monoclonal) |
BioLegend |
Cat# 339808 |
Flow cytometry |
| RRID:AB_10895898 |
| Anti-PD-L1-BV-785 (clone 10F.9G2, rat, monoclonal) |
BioLegend |
Cat# 124331 |
Flow cytometry |
| RRID:AB_2629659 |
| Recombinant proteins |
| Recombinant murine G-CSF |
PeproTech |
Cat# 250-05 |
G-CSF treatment |
| 10 μg, daily injection |
| Recombinant murine TNF |
PeproTech |
Cat# 315-01A |
TNF stimulation |
| 10 ng/ml, 4 hours |
| Viral vector |
| Ad5CMVFlpo (adenoviral vector) |
|
|
University of Iowa Viral Vector Core Facility |
| Sequence-based reagents |
|
Rpl30 (Mus musculus) |
Thermo Fisher Scientific |
Mm01611464_g1 |
TaqMan probe |
| (Housekeeping gene) |
| Slc2a1 (Glut1) (Mus musculus) |
Thermo Fisher Scientific |
Mm00441480_m1 |
TaqMan probe |
| Slc2a3 (Glut3) (Mus musculus) |
Thermo Fisher Scientific |
Mm00441483_m1 |
TaqMan probe |
|
Arg1 (Mus musculus) |
Thermo Fisher Scientific |
Mm00475988_m1 |
TaqMan probe |
|
Hk2 (Mus musculus) |
Thermo Fisher Scientific |
Mm00443385_m1 |
TaqMan probe |
|
Siglecf (Siglec5) (Mus musculus) |
Thermo Fisher Scientific |
Mm00523987_m1 |
TaqMan probe |
|
Ikbke (Mus musculus) |
Thermo Fisher Scientific |
Mm00444862_m1 |
TaqMan probe |
|
Apoe (Mus musculus) |
Thermo Fisher Scientific |
Mm01307193_g1 |
TaqMan probe |
|
Pfkp (Mus musculus) |
Thermo Fisher Scientific |
Mm00444792_m1 |
TaqMan probe |
|
Cxcl3 (Mus musculus) |
Thermo Fisher Scientific |
Mm01701838_m1 |
TaqMan probe |
|
Grx1 (Mus musculus) |
Thermo Fisher Scientific |
Mm01352826_g1 |
TaqMan probe |
|
Il23a (Mus musculus) |
Thermo Fisher Scientific |
Mm00518984_m1 |
TaqMan probe |
|
Padi4 (Mus musculus) |
Thermo Fisher Scientific |
Mm01341658_m1 |
TaqMan probe |
|
Csf3 (G-CSF) (Mus musculus) |
Thermo Fisher Scientific |
Mm00438334_m1 |
TaqMan probe |
| Chemicals |
| Isoflurane |
Piramal |
Cat# 56.761.002 |
Anaesthetic |
| BrdU |
Merck |
Cat# 10280879001 |
BrdU assay |
| 2 mg of freshly prepared BrdU into 100 μl of PBS 1X |
| Glucose |
VWR |
Cat# 1.08337.0250 |
Seahorse |
| 10 mM glucose |
| 2-deoxyglucose (2-DG) |
Merck |
Cat# D8375-5G |
Seahorse |
| 50 mM 2-DG |
| Crystal violet |
Merck |
Cat# C3886 |
|
| Experimental Models: Cell lines |
| B16-F1 (Mus musculus) |
|
RRID:CVCL_0158 |
Melanoma lung metastases |
| SV2 (Mus musculus) |
|
|
Developed in our laboratory |
| Experimental Models: Organism/Strains |
|
KrasLox-STOP-Lox-G12D/WT (K) |
The Jackson Laboratory |
RRID:IMSR_JAX:008179 |
KrasLoxSTOPLox-G12D/WT
|
| K and P were interbred at EPFL |
|
Tp53Flox/Flox (P) |
The Jackson Laboratory |
RRID:IMSR_JAX:008462 |
Trp53Flox/Flox
|
| K and P were interbred at EPFL |
|
KrasFrt-STOP-Frt-G12D/WT (KFrt) |
The Jackson Laboratory |
RRID:IMSR_JAX:008653 |
KPFrt were provided by D.G. Kirsch, Duke University Medical Center in a mixed 129-C57BL/6 background |
|
Tp53Frt/Frt (PFrt) |
The Jackson Laboratory |
RRID:IMSR_JAX:017767 |
KPFrt were provided by D.G. Kirsch, Duke University Medical Center in a mixed 129-C57BL/6 background |
|
Glut1Flox/Flox (G1) |
The Jackson Laboratory |
RRID:IMSR_JAX:031871 |
Glut1Flox/Flox were provided by E. Dale Abel, University of Iowa in a C57BL/6 background |
|
Glut3Flox/Flox (G3) |
|
|
Duke University Transgenic Facility |
|
Ly6gCre (Catchup) |
|
|
Ly6gCre (Catchup) were provided by M. Gunzer, University Duisburg-Essen, in a C57BL/6 background |
|
KrasFrt-STOP-Frt-G12D/WT; Tp53Frt/Frt; Ly6gCre/WT; Glut1Flox/Flox
|
|
|
G1, Ly6gCre and KPFrt mice were interbred at EPFL |
|
KrasFrt-STOP-Frt-G12D/WT; Tp53Frt/Frt; Ly6gWT/WT; Glut1Flox/Flox
|
|
|
G1, Ly6gCre and KPFrt mice were interbred at EPFL |
|
KrasFrt-STOP-Frt-G12D/WT; Tp53Frt/Frt; Ly6gCre/WT; Glut1WT/WT
|
|
|
G1, Ly6gCre and KPFrt mice were interbred at EPFL |
| Commercial assays and kits |
| Anti-mouse CD45 MicroBeads (clone 30F11.1, monoclonal) |
Miltenyi Biotec |
Cat# 130-052-301 |
CD45 magnetic cell sorting |
| RRID:AB_2877061) |
| Anti-mouse Ly-6G MicroBead (monoclonal) |
Miltenyi Biotec |
Cat# 130-092-332 |
Ly-6G magnetic cell sorting |
| Anti-mouse Ly-6G MicroBeads UltraPure (clone REA526, monoclonal) |
Miltenyi Biotec |
Cat# 130-120-337 |
Ly-6G magnetic cell sorting |
| Centricon Plus-70 Centrifugal Filter (3 kDa cutoff) |
Merck |
Cat# UFC700308 |
Size exclusion separation of proteins |
| High-Capacity cDNA Reverse Transcription Kit |
Thermo Fisher Scientific |
Cat# 4368814 |
Reverse transcription |
| BCA Protein Assay Kit |
Merck |
Cat# 71285-3 |
Protein quantification |
| CountBright™ Absolute Counting Beads |
Thermo Fisher Scientific |
Cat# C36950
|
Flow cytometry |
| Anti-rabbit Immpress HRP (horse) |
Vector Laboratories |
Cat# MP-7401 |
Peroxidase Polymer Detection Kit |
| RRID:AB_2336529 |
| Rhodamine-6G |
Roche Diagnostics |
Cat# 07988168001 |
TSA kit |
| DCC |
Roche Diagnostics |
Cat# 07988192001 |
TSA kit |
| Red 610 |
Roche Diagnostics |
Cat# 07988176001 |
TSA kit |
| Cyanine 5 |
Roche Diagnostics |
Cat# 07551215001 |
TSA kit |
| Glucose-uptake cell-based assay kit |
Cayman Chemical |
Cat# CAY-600470-1 |
Glucose Uptake Cell-Based Assay Kit |
| CellTiter-Glo® Luminescent Cell Viability Assay |
Promega |
Cat# G7570 |
ATP production |
| Software and Algorithms |
| Analyze 12.0 |
PerkinElmer |
|
Tumor volume analysis |
| OsiriX MD |
Pixmeo |
RRID:SCR_013618 |
Tumor volume analysis |
| QuPath |
|
RRID:SCR_018257 |
|
| Fiji |
|
RRID:SCR_002285 |
|
| FlowJo |
|
RRID:SCR_008520 |
Flow cytometry |
| GraphPad Prism |
|
RRID:SCR_002798 |
|
| (version 6) |
| Instruments |
| X-Ray microtomography |
PerkinElmer |
|
Quantum FX microCT |
| 20 mm2 collimator |
|
|
Radiotherapy treatment |
| 13 mA, 3 mm Cu filter, 225 keV |
| 11.6 Gy with a single dose in 256 seconds |
| Automated Ventana Discovery ULTRA |
Roche Diagnostics |
|
Human sample multiplexing |
| Olympus slide scanner |
Olympus |
VS120-L100 |
Slide scanner |
| XF96 Extracellular Flux analyser |
Seahorse Bioscience |
XF96 |
Seahorse |
| LSRII SORP |
Becton Dickinson |
|
Flow cytometry |
