Genetic reagent (M. musculus) | Cd4Cre | PMID: 27345256, PMID: 11728338 | | |
Genetic reagent (M. musculus) | Cd4Cre+ Mycfl/fl | PMID: 19423665, PMID: 19342639, PMID: 11742404 | | |
Genetic reagent (M. musculus) | Cd4Cre+ Slc7a5fl/fl | PMID: 23525088, PMID: 24586861 | | |
Genetic reagent (M. musculus) | GFP-MycKI | PMID: 18196519, PMID: 26136212, PMID: 23021216 | | |
Genetic reagent (M. musculus) | OT1 | PMID: 8287475 | | maintained in house as an OT1 TCR transgene heterozygote on a CD45.1 (Ly5.1) background |
Antibody | Anti-CD3 (armenian hamster, monoclonal) | Thermo Fisher Scientific | Cat # 14-0031-82, RRID:AB_467049 | T cell activation: 0.5 or 1 μg/ml as indicated in Materials and methods |
Antibody | Anti-CD28 (syrian hamster, monoclonal) | Thermo Fisher Scientific | Cat # 16-0281-82, RRID:AB_468921 | T cell activation: 0.5 or 3 μg/ml as indicated in Materials and methods |
Antibody | Anti-CD4 (rat, monoclonal) | BD Biosciences | Cat # 553650, RRID:AB_394970; Cat# 552775, RRID:AB_394461; Cat# 553047, RRID:AB_394583 | cell surface staining 1:200 |
Antibody | Anti-CD4 (rat, monoclonal) | Thermo Fisher Scientific | Cat# 47-0042-82, RRID:AB_1272183 | cell surface staining 1:200 |
Antibody | Anti-CD8a (rat, monoclonal) | Biolegend | Cat# 100708, RRID:AB_312747; Cat# 100722, RRID:AB_312761 Cat# 100738, RRID:AB_11204079 | cell surface staining 1:200 |
Antibody | Anti-CD8a (rat, monoclonal) | BD Biosciences | Cat# 551162, RRID:AB_394081 | cell surface staining 1:200 |
Antibody | Anti-CD69 (armenian hamster, monoclonal) | ThermoFisher Scientific | Cat# 17-0691-82, RRID:AB_1210795 | cell surface staining 1:200 |
Antibody | Anti-CD69 (armenian hamster, monoclonal) | Biolegend | Cat# 104514, RRID:AB_492843 | cell surface staining 1:200 |
Antibody | Anti-CD69 (armenian hamster, monoclonal) | BD Biosciences | Cat# 553237, RRID:AB_394726 | cell surface staining 1:200 |
Antibody | Anti-B220 (rat, monoclonal) | BD Biosciences | Cat# 553087, RRID:AB_394617 | cell surface staining 1:200 |
Antibody | Anti-NK1.1 (mouse, monoclonal) | Biolegend | Cat# 108706, RRID:AB_313393 | cell surface staining 1:200 |
Antibody | Anti-CD11b (rat, monoclonal) | Biolegend | Cat# 101206, RRID:AB_312789 | cell surface staining 1:200 |
Antibody | Anti-CD25 (rat, monoclonal) | BD Biosciences | Cat# 553072, RRID:AB_394604 | cell surface staining 1:200 |
Antibody | Anti-CD62L (rat, monoclonal) | Thermo Fisher Scientific | Cat# 12-0621-83, RRID:AB_465722 | cell surface staining 1:200 |
Antibody | Anti-TCRb (armenian hamster, monoclonal) | Thermo Fisher Scientific | Cat# 45-5961-82, RRID:AB_925763 | cell surface staining 1:200 |
Antibody | Anti-CD44 (rat, monoclonal) | BD Biosciences | Cat# 559250, RRID:AB_398661 | cell surface staining 1:200 |
Antibody | Anti-Thy1.2 (rat, monoclonal) | BD Biosciences | Cat# 553006, RRID:AB_394545 | cell surface staining 1:200 |
Antibody | Anti-CD45.1 (mouse, monoclonal) | Biolegend | Cat # 110714, RRID:AB_313503 | cell surface staining 1:200 |
Antibody | Anti-CD45.2 (mouse, monoclonal) | Biolegend | Cat # 109816, RRID:AB_492868 | cell surface staining 1:200 |
Antibody | Anti-mouse CD16/CD32 Fc Block, (rat, monoclonal) | BD Biosciences | Cat # 553141, RRID:AB_394656 | Fc block 1:100 |
Antibody | c-Myc (D84C12) XP (rabbit, monoclonal) | Cell Signaling Technologies | Cat# 5605, RRID:AB_1903938 | intracellular staining 1:200 |
Antibody | Anti-rabbit A647 (goat) | Cell Signaling Technologies | Cat # 4414, RRID:AB_10693544 | intracellular staining 1:1000 |
Antibody | Anti-IFNg (rat, monoclonal) | Biolegend | Cat # 505810, RRID:AB_315404 | intracellular cytokine staining 1:100 |
Antibody | Anti-Granzyme B | Thermo Fisher Scientific | Cat# 17-8898-82, RRID:AB_2688068 | intracellular cytokine staining 1:200 |
Chemical compound, drug | DAPI | Thermo Fisher Scientific | D1306 | 1 µg/mL |
Chemical compound, drug | Kynurenine | Sigma | Cat# K8625 | 200 µM |
Chemical compound, drug | BCH | Sigma | Cat# A7902 | 10 mM |
Chemical compound, drug | Rapamycin | Merck/Calbiochem | Cat# 553211 | 20 nM |
Commercial assay or kit | Rneasy minikit | Qiagen | Cat # 74104 | |
Commercial assay or kit | iScript cDNA Synthesis kit | Biorad | Cat#1708891 | |
Commercial assay or kit | iTaq Universal SYBRGreen Supermix | Biorad | Cat# 1725121 | |
Commercial assay or kit | EZQ protein quantitation kit | Thermo Fisher Scientific | R33200 | |
Commercial assay or kit | Sera-Mag SpeedBead Carboxylate-modified magnetic particles (hydrophilic) | GE Lifesciences | cat# 45152105050250 | |
Commercial assay or kit | Sera-Mag SpeedBead Carboxylate-modified magnetic particles (hydrophobic) | GE Lifesciences | cat# 65152105050250 | |
Commercial assay or kit | CBQCA protein quantitation kit | Thermo Fisher Scientific | C6667 | |
Commercial assay or kit | HiPPR Detergent Removal Spin Column Kit | Thermo Fisher Scientific | Cat# 88305 | |
Commercial assay or kit | EasySep CD8 T cell isolation kit | STEMCELL Technologies, UK | Cat # 19853 | |
Commercial assay or kit | Golgi Plug | BD Biosciences | Cat# 555029 | |
Commercial assay or kit | eBioscience Intracellular Fixation and Permeabilization Buffer Set | Thermo Fisher Scientific | Cat# 88-8824-00 | |
Commercial assay or kit | CFSE | Thermo Fisher Scientific/Invitrogen | Cat# C34554 | 5 µM |
Peptide, recombinant protein | IL7 | Peprotech | Cat# 217–17 | 5 ng/mL |
Peptide, recombinant protein | IL2 | Novartis, UK | Proleukin | 20 ng/ml |
Peptide, recombinant protein | IL12 | Peprotech | Cat#210–12 | 2 ng/ml |
Sequence based reagent | Slc7a5 forward primer | | | AAG GCT GCG ACC CGT GTG |
Sequence based reagent | Slc7a5 reverse primer | | | ATC ACC TTG TCC CAT GTC CTT CC |
Sequence based reagent | Slc7a1 forward primer | | | GGA GCT TTG GC CTT CAT CAC T |
Sequence based reagent | Slc7a1 reverse primer | | | CAG CAC CCC AGG AGC ATT CA |
Sequence based reagent | Slc1a5 forward primer | | | GCC ATC ACC TCC ATC AAC GAC T |
Sequence based reagent | Slc1a5 reverse primer | | | AGA GCG GAA GGC AGC AGA CAC |
Sequence based reagent | TBP forward primer | | | GTG AAT CTT GGC TGT AAA CTT GAC CT |
Sequence based reagent | TBP reverse primer | | | CGC AGT TGT CCG TGG CTC T |
Software, algorithm | FlowJo software | Treestar | | versions 9 and 10 |
Software, algorithm | Maxquant | https://www.maxquant.org, PMID: 19029910 | | version 1.6.2.6 |
Software, algorithm | Perseus | https://www.maxquant.org/perseus, PMID: 27348712 | | version 1.6.6.0 |
Other | RPMI 1640 | Thermo Fisher Scientific/GIBCO | Cat# 21875–034 | |
Other | RPMI - glutamine | Thermo Fisher Scientific/GIBCO | Cat# 42401–018 | |
Other | RPMI - methionine | DC Biosciences Ltd | | custom made RPMI without methionine and arginine - supplemented back the arginine (0.2 g/L) to RPMI levels |
Other | RPMI - leucine | Sigma | Cat# R1780 SAFC | supplemented back arginine (0.2 g/L) and lysine (0.04 g/L)to RPMI levels |
Other | HBSS | Thermo Fisher Scientific/GIBCO | Cat# 14025–050 | used this as amino acid-free media |
Other | FBS | Thermo Fisher Scientific/GIBCO | Cat # 10270106 | |
Other | FBS, dialyzed | Thermo Fisher Scientific/GIBCO | Cat# 26400044 | |
Other | Arginine | Sigma | Cat# A5006 | |
Other | Lysine | Sigma | Cat# L5501 | |