| Antibodies |
| anti-mouse CD4 PE-Cy7 (clone RM4-5) |
Biolegend |
Cat # 100528 |
| anti-mouse IFNγ PE-CF594 (clone XMG1.2) |
BD Biosciences |
Cat # 562303 |
| anti-mouse IL17 PE (clone TC11-18H10) |
BD Biosciences |
Cat # 559502 |
| anti-mouse CD44 PE (clone IM7) |
Thermo Fisher |
Cat # 12-0441-83 |
| anti-mouse Ly-6G, PE (clone 1A8) |
BD Biosciences |
Cat # 551461 |
| anti-mouse Siglec-F, APC (clone 1RNM44N) |
eBioscience |
Cat # 50-1702-82 |
| anti-mouse IL-4, APC (clone 11B11) |
BD Biosciences |
Cat # 557739 |
| anti-mouse IL-13, PE (clone eBio13A) |
eBioscience |
Cat # 12-7133-82 |
| anti-mouse IL-17, FITC (clone TC11-18H10) |
BD Biosciences |
Cat # 560220 |
| anti-mouse IL-5, PE (clone TRFK5) |
BD Biosciences |
Cat # 554395 |
| anti-mouse IL-10, APC (clone JES5-16E3) |
BD Biosciences |
Cat # 554468 |
| anti-mouse IL-2, FITC (clone JES6-5H4) |
BD Biosciences |
Cat # 554427 |
| anti-mouse CD62L, PE (clone MEL-14) |
BD Biosciences |
Cat #553151 |
| anti-mouse CD8, APC (clone 53-6.7) |
BD Biosciences |
Cat #553035 |
| anti-mouse CD4, BV421 (clone RM4-5) |
Biolegend |
Cat # 100563 |
| anti-mouse CD25, PE-cy5 (clone PC61.5) |
eBioscience |
Cat # 15-0251-82 |
| anti-mouse ICOS, APC (clone C398.4A) |
BD Pharmingen |
Cat # 565882 |
| anti-mouse Nrp1, PE (clone 3E12) |
Biolegend |
Cat # 145204 |
| anti-mouse CD44, BV510 (clone IM7) |
BD Biosciences |
Cat # 563114 |
| anti-mouse GITR, PE-cy7 (clone DTA-1) |
BD Biosciences |
Cat # 558140 |
| anti-mouse PD1, APC (clone J43) |
Invitrogen |
Cat # 17-9985-82 |
| anti-mouse CTLA4, PE-cy7 (clone UC10-4B9) |
Biolegend |
Cat # 106314 |
| anti-mouse Foxp3, FITC (clone FJK-16s) |
Invitrogen |
Cat # 11-5773-82 |
| anti-mouse Tbet, APC (clone eBio4B10) |
eBioscience |
Cat # 50-5825-82 |
| anti-mouse Gata3, PE (clone TWAJ) |
eBioscience |
Cat # 12-9966-42 |
| anti-mouse RORγ, PE-TR (clone B20) |
Invitrogen |
Cat # 61-6981-82 |
| anti-mouse Ki67, PE-cy7 (clone SolA15) |
Invitrogen |
Cat # 25-5698-82 |
| anti-Rictor (clone 53A2) |
CST |
Cat # 2114 |
| anti-Raptor (24C12) |
CST |
Cat # 2280 |
| anti-Beta actin |
CST |
Cat # 4967 |
| anti-pAKT (S473) (clone D9E) |
CST |
Cat # 4060 |
| anti-pAKT (T308) (clone C31E5E) |
CST |
Cat # 2965 |
| anti-AKT (clone C67E7) |
CST |
Cat # 4691 |
| anti-pFoxo1a (S256) |
CST |
Cat # 9461 |
| anti-Foxo1a (clone C29H4) |
CST |
Cat # 2880 |
| anti-pS6 (clone D57.2.2E) |
CST |
Cat # 4858 |
|
|
|
| Chemicals, Peptides, and Recombinant Proteins |
| Cockroach antigens |
Greer Laboratory |
Item# B26 |
| Aluminum hydroxide (Alum) |
MilliporeSigma |
Cat # 239186 |
| Anti-mouse CD3 mAb (clone 2C11) |
Envigo Bioproducts |
N/A |
| Transforming growth factor β (TGF-β) |
PeproTech |
Cat# 100-21 |
| Diphtheria toxin (DTx) |
Sigma-Aldrich |
Cat # D0564 |
| Pertussis toxin (PTx) |
Sigma-Aldrich |
Cat # P2980 |
| Dexamethasone-Water Soluble (Dex) |
Sigma-Aldrich |
Cat # D2915 |
| MOG (35–55) |
Biosynthesis |
Cat # 12668-01 |
| PMA (phorbol 12-myristate 13-acetate) |
Sigma-Aldrich |
Cat # P1585 |
| Ionomycin |
Sigma-Aldrich |
Cat # I9657 |
| TRIzol |
Invitrogen |
Cat # 15596026 |
| Power SYBR Green PCR Master Mix |
Thermo Fisher |
Cat # 4367659 |
|
|
|
| Recombinant DNA |
| Myc_Rictor |
Addgene |
Plasmid #11367 |
|
|
|
| Critical Commercial Assays |
| Mouse IL-10 DuoSet ELISA |
R&D systems |
Cat # DY417-05 |
| Mouse IFN-gamma DuoSet ELISA |
R&D systems |
Cat # DY485-05 |
| miScript II RT Kit |
Qiagen |
Cat # 218161 |
| miScript SYBR Green PCR Kit |
Qiagen |
Cat # 218073 |
| Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit |
BD Biosciences |
Cat # 560485 |
| CBA mouse IL-13 Enhanced Sensitivity Flex Set |
BD Biosciences |
Cat # 562273 |
| Quick-RNA™ Miniprep Kit |
Zymo research |
Cat # R1051 |
| Mouse T cell nucleofector kit |
Amaxa |
Cat # VPA-1006 |
| MojoSort™ Mouse CD4 Naïve T Cell Isolation Kit |
Biolegend |
Cat # 480040 |
| SuperSignal West Femto Maximum Sensitivity Substrate |
Thermo Fisher |
Cat # 34095 |
| Mouse: GAPDH FAM/MGB probe |
Thermo Fisher |
ID# 4352661 |
| Mouse: Foxp3 FAM/MGB probe |
Thermo Fisher |
ID# Mm00475162_m1 |
| Mouse: IFNg FAM/MGB probe |
Thermo Fisher |
ID# Mm01168134_m1 |
| Mouse: IL17 FAM/MGB probe |
Thermo Fisher |
ID# Mm00439618_m1 |
| Mouse: Rragd FAM/MGB probe |
Thermo Fisher |
ID# Mm00546741_m1 |
| Mouse: Galnt9 FAM/MGB probe |
Thermo Fisher |
ID# Mm01196716_m1 |
| Mouse: Esam FAM/MGB probe |
Thermo Fisher |
ID# Mm00518378_m1 |
| Mouse: Evl FAM/MGB probe |
Thermo Fisher |
ID# Mm00468405_m1 |
| Mouse: Nabp FAM/MGB probe |
Thermo Fisher |
ID# Mm00658947_m1 |
| Mouse: Rictor FAM/MGB probe |
Thermo Fisher |
ID# Mm01307318_m1 |
| Mouse: Raptor FAM/MGB probe |
Thermo Fisher |
ID# Mm01242613_m1 |
| Mouse: Dusp1 FAM/MGB probe |
Thermo Fisher |
ID# Mm00457274_g1 |
| Mouse: Fkbp5 FAM/MGB probe |
Thermo Fisher |
ID# Mm00487406_m1 |
| Mouse: Muc5ac FAM/MGB probe |
Thermo Fisher |
ID# Mm01276718_m1 |
| Mouse: Muc5b FAM/MGB probe |
Thermo Fisher |
ID# Mm00466391_m1 |
| Mouse: Hk2 FAM/MGB probe |
Thermo Fisher |
ID# Mm00443385_m1 |
| Mouse: Hif1a FAM/MGB probe |
Thermo Fisher |
ID# Mm00468869_m1 |
| Mm_miR-15b_2 miScript Primer Assay |
Qiagen |
ID# MS00011235 |
| Mm_miR-16_2 miScript Primer Assay |
Qiagen |
ID# MS00037366 |
| Mm_miR-146_1 miScript Primer Assay |
Qiagen |
ID# MS00001638 |
| Mm_miR-155_1 miScript Primer Assay |
Qiagen |
ID# MS00001701 |
| Mm_miR-342_1 miScript Primer Assay |
Qiagen |
ID# MS00002184 |
| 5 nmol IDT miRNA Inhibitor_NC5; mG/ZEN/mCmGmAmCmUmAmUmAmCmGmCmGmCmAmAmUmAmUmGmG/3ZEN/ |
IDT |
Ref # 210223534 |
| 5 nmol IDT miRNA Inhibitor_mmu-miR342-3p; mA/ZEN/mCmGmGmGmUmGmCmGmAmUmUmUmCmUmGmUmGmUmGmAmG/3ZEN/ |
IDT |
Ref # 210223535 |
| mirVan miRNA Mimic, Negative Control #1 |
Thermo Fisher |
Cat # 4464058 |
| mirVana miR-342 Mimic |
Thermo Fisher |
ID#MC12328 |
| Accell Mouse Rictor siRNA |
Horizon Discovery |
Cat # A06459813-0005 |
| Seahorse XF Glycolysis Stress Test Kit |
Agilent |
Cat # 103020-100 |
| Seahorse XF Cell Mito Stress Test Kit |
Agilent |
Cat # 103015-100 |
|
|
|
| Deposited Data |
| RNAseq |
This paper |
GEO: GSE136969
|
|
|
|
| Experimental Models: Organisms/Strains |
| C57BL/6 mice (B6) |
Jackson Laboratory |
JAX: 000664 |
| B6 Foxp3GFP (B6 background) |
Generated by V. Kuchroo and Provided by Y. Belkaid (Bettelli et al., 2006) |
N/A |
| B6.129(Cg)-Foxp3tm3(DTR/GFP)Ayr/J (B6 background) |
Jackson Laboratory |
JAX: 016958 |
| B6.Cg-Nr3c1tm1.1Jda/J (B6 background) |
Jackson Laboratory |
JAX: 021021 |
| B6.129(Cg)-Foxp3tm4(YFP/icre)Ayr/J (B6 background) |
Jackson Laboratory |
JAX: 016959 |
|
|
|
| Oligonucleotides |
| GAPDH forward: 5’- ATGCCTGCTTCACCACCTTCT-3’ |
IDT |
N/A |
| GAPDH reverse: 5’- CATGGCCTTCCGTGTTCCTA-3’ |
IDT |
N/A |
| common GR forward: 5’-AAAGAGCTAGGAAAAGCCATTGTC-3’ |
IDT |
N/A |
| GRα reverse: 5’-TCAGCTAACATCTCTGGGAATTCA-3’ |
IDT |
N/A |
| Gilz forward: 5’- AATGCGGCCACGGATG -3’ |
IDT |
N/A |
| Gilz reverse: 5’- GGACTTCACGTTTCAGTGGACA -3’ |
IDT |
N/A |
|
|
|
| Software and Algorithms |
| Prism 7 |
GraphPad |
https://www.graphpad.com/scientific-software/prism/ |
| Flowjo |
TreeSta |
https://www.flowjo.com/solutions/flowjo/downloads |
| Seahorse Wave |
Seahorse Wave |
|
|
|
|
