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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Cell Metab. 2020 Nov 17;32(6):996–1011.e7. doi: 10.1016/j.cmet.2020.10.022

KEY RESOURCES TABLE

REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-CD4 (RM4–5) Thermo Fisher Scientific Cat# 48–0042-80; RRID:AB_1272231
Anti-CD4 (RM4–4, in vivo labeling) BioLegend Cat# 116006; RRID:AB_313691
Anti-CD8α (53–6.7) BioLegend Cat# 100742; RRID:AB_2563056
Anti-TCRβ (H57–597) Thermo Fisher Scientific Cat# 47–5961-82; RRID:AB_1272173
Anti-CD44 (1M7) BioLegend Cat# 103047; RRID:AB_2562451
Anti-CD62L (MEL-14) Tonbo Biosciences Cat# 60–0621; RRID:AB_469632
Anti-CD69 (H1.2F3) Thermo Fisher Scientific Cat# 48–0691-82; RRID:AB_10719430
Anti-PD-1 (J43) Thermo Fisher Scientific Cat# 17–9985-80; RRID:AB_11149860
Anti-CD45.1 (A20) BioLegend Cat# 110736; RRID:AB_2562564
Anti-CD45.2 (104) Thermo Fisher Scientific Cat# 48–0454-80; RRID:AB_11039533
Anti-CD25 (PC61.5) Thermo Fisher Scientific Cat# 17–0251-82; RRID:AB_469366
Anti-Foxp3 (FJK-16s) Thermo Fisher Scientific Cat# 12–5773-80; RRID:AB_465935
Anti-Ki67 (SolA15) Thermo Fisher Scientific Cat# 25–5698-80; RRID:AB_11217689
Anti-Bim (C34C5) Cell Signaling Technology Cat# 2933; RRID:AB_1030947
Anti-GFP-Alexa Fluor 488 (FM264G) BioLegend Cat# 338008; RRID:AB_2563288
Anti-IFN-γ (XMG1.2) Thermo Fisher Scientific Cat# 17–7311-81; RRID:AB_469503
Anti-IL-4 (11B11) Thermo Fisher Scientific Cat# 12–7041-71; RRID:AB_466154
Anti-IL-17 (eBio17B7) Thermo Fisher Scientific Cat# 45–7177-80; RRID:AB_925754
Anti-CD3 (145–2C11) Bio-X-Cell Cat# BE0001; RRID:AB_2714218
Anti-CD28 (37.51) Bio-X-Cell Cat# BE0015–1; RRID:AB_1107624
Anti-β-actin (C4)-HRP Santa Cruz Biotechnology Cat# sc-47778 HRP; AB_2714189
Anti-β-tubulin Cell Signaling Technology Cat# 2146, RRID:AB_2210545
Anti-phospho-S6 (S235/236) (D57.2.2E) Cell Signaling Technology Cat# 4858; RRID:AB_916156
Anti-phospho-4EBP1 (Thr37/46) (236B4)
Cell Signaling Technology Cat# 2855; RRID: AB_560835
Anti-phospho-4EBP1 (Thr37/46) (236B4)
-Alexa Fluor 647 conjugate
Cell Signaling Technology Cat# 5123
RRID: AB_2097838
Anti-phospho-S6 (S235/236) (D57.2.2E)-Pacific Blue™ conjugate Cell Signaling Technology Cat# 8520, RRID:AB_2797646
Anti-Mcl1 (LVUBKM) Thermo Fisher Scientific  Cat# 12–9047-42, RRID:AB_2762598
Anti-Bcl2 (BCL/10C4) BioLegend Cat# 633510, RRID:AB_2274702
Anti-ICOS (7E.17G9) Thermo Fisher Scientific Cat# 12–9942-82, RRID:AB_466274
Anti-CTLA4 (UC10–4B9) BioLegend Cat# 106312, RRID:AB_2563063
Anti-KLRG1 (2F1) BioLegend Cat# 138419, RRID:AB_2563357
Anti-CXCR3 (CXCR3–173) BioLegend Cat# 126531, RRID:AB_2563160
Anti-CD73 (TY/11.8) BioLegend Cat# 127212, RRID:AB_11219190
Anti-CD5 (53–7.3) Thermo Fisher Scientific Cat# 12–0051-81, RRID:AB_465522
Anti-Bim (C34C5) Cell Signaling Technology Cat# 12186, RRID:AB_2797842
Anti-Rap1A (C-17) Santa Cruz Biotechnology Cat# sc-1482, RRID:AB_2177124
Anti-HDJ2 (KA2A5.6) Lab Vision Cat# MS-225-P0, RRID:AB_61951
Anti-Rabbit IgG HRP conjugate Promega Cat# W4011, RRID:AB_430833
Anti-Mouse IgG HRP conjugate Promega Cat# W4021, RRID:AB_430834
Anti-Goat IgG HRP conjugate (sc-2345) Santa Cruz Biotechnology Cat# sc-2354, RRID:AB_628490
Chemicals, Peptides, and Recombinant Proteins
7-Aminoactinoymycin D (7AAD) Sigma-Aldrich Cat# A9400
Simvastatin Selleck Chemicals Cat# S1792
FTI-277 Tocris Cat# 2407
GGTI-2147 Sigma-Aldrich Cat# 191102–87-1
(+)-Mevalonolactone (dehydrated mevalonate) Sigma-Aldrich Cat# M4667–5G
Farnesyl pyrophosphate ammonium salt (FPP) Sigma-Aldrich Cat# 13058–04-3
Geranylgeranyl pyrophosphate ammonium salt (GGPP) Sigma-Aldrich Cat# G6025
Fixable Viability Dye eFluor™ 780 Thermo Fisher Scientific Cat# 65–0865-18
Recombinant human IL-2 (in vitro stimulation) R&D Systems Cat# 202-IL
Phobol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat# P8139
Ionomycin Sigma-Aldrich Cat# I0634
Golgi-STOP BD Biosciences Cat# 554724
Recombinant human IL-6 National Cancer Institute Cat# NSC720836
Recombinant murine IL-3 Thermo Fisher Scientific Cat# PMC0034
Recombinant murine stem-cell factor (SCF) Thermo Fisher Scientific Cat# PMC2115
ABI probe: Pggt1b Thermo Fisher Scientific Mm01266356_m1
ABI probe: Icos Thermo Fisher Scientific Mm00497600_m1
ABI probe: Actb Thermo Fisher Scientific Mm00607939_s1
Commercial Reagents
Foxp3 transcription factor staining buffer set Thermo Fisher Scientific Cat# 00–5523-00
Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit BD Biosciences Cat# 554714
CD4 (3LT4) Microbeads, mouse Miltenyi Biotec Cat# 130–117-043
Lineage cell depletion kit Miltenyi Biotec Cat# 130–090-858
Transfection reagent Mirus Cat# MIR2706
10× Perm/Wash Buffer BD Biosciences Cat# 554723
Clariom S mouse array Thermo Fisher Scientific Cat# 902930
BD Phosflow™ Fix Buffer I BD Biosciences Cat# 557870
Phosflow™ Perm Buffer III BD Biosciences Cat# 558050
CellTrace™ Violet Cell Proliferation Kit Thermo Fisher Scientific Ca# C34557
Deposited Data
Fntb- or Pggt1b-deficient Treg-cell microarray data from mixed bone marrow chimeras GEO (http://www.ncbi.nlm.nih.gov/geo/) GSE158863
In vitro cTreg-cell microarray data from DMSO, FTI, or GGTI treatments GEO (http://www.ncbi.nlm.nih.gov/geo/) GSE158863
ATAC-Seq data GEO (http://www.ncbi.nlm.nih.gov/geo/) GSE158863
Experimental Models: Cell Lines
Plat-E Dr. Yun-Cai Liu, La Jolla Institute of Immunology N/A
Experimental Models: Organisms/Strains
Mouse: C57BL/6J The Jackson Laboratory Cat# 000664; RRID:IMSR_JAX:000664
Mouse: Fntbfl/fl M. Bergo (Liu et al., 2010) N/A
Mouse: Pggt1bfl/fl M. Bergo (Khan et al., 2011) N/A
Mouse: Hmgcrfl/fl A. Waisman N/A
Mouse: CD45.1+: B6.SJL-Ptprca Pep3b/BoyJ The Jackson Laboratory Cat# 002014; RRID:IMSR_JAX:002014
Mouse: Cd4 Cre: Tg(Cd4-cre)1Cwi/BfluJ The Jackson Laboratory Cat# 017336; RRID:IMSR_JAX:017336
Mouse: Rag1−/−: Rag1tm1Mom/J The Jackson Laboratory Cat# 002216; RRID:IMSR_JAX:002216
Mouse: Foxp3YFP-Cre A. Rudensky (Rubtsov et al., 2008) N/A
Mouse: Rac1fl/fl The Jackson Laboratory Cat# JAX:005550, RRID:IMSR_JAX:005550
Mouse: Rac2−/− The Jackson Laboratory Cat# JAX:004197, RRID:IMSR_JAX:004197
Mouse: Tsc1−/− The Jackson Laboratory Cat# JAX:005680, RRID:IMSR_JAX:005680
Mouse: C57BL/6-Tg(Nr4a1-EGFP/cre)820Khog/J (also known as Nur77-GFP) The Jackson Laboratory Cat# JAX:016617, RRID:IMSR_JAX:016617
Mouse: C57BL/6-Foxp3tm1Flv/J (also known as Foxp3-RFP) The Jackson Laboratory Cat# JAX: 008374
RRID:IMSR JAX: 008374
Oligonucleotides
Fntb-F: 5’-AACCGCTGTATAGTCTGAGACC-3’ This paper N/A
Fntb-R: 5’-TCTCCCTCTGCAAAATGAGCC-3’ This paper N/A
Hmgcr-F: 5’-AGCTTGCCCGAATTGTATGTG-3’ (Zeng et al., 2013) N/A
Hmgcr-R: 5’-TCTGTTGTGAACCATGTGACTTC-3’ (Zeng et al., 2013) N/A
Hmgcs1-F: 5’-AACTGGTGCAGAAATCTCTAGC-3’ (Zeng et al., 2013) N/A
Hmgcs1-R: 5’-GGTTGAATAGCTCAGAACTAGCC-3’ (Zeng et al., 2013) N/A
Fnta-F: 5’- CCCTATGGACGACGGGTTTC-3’ This paper N/A
Fnta-R:5’- TGATCTGGACCACTGGGTTAG-3’ This paper N/A
Actb-F: 5’-GACAGGATGCAGAAGGAGATTACTG-3’ This paper N/A
Actb-R: 5’-GCTGATCCACATCTGCTGGAA-3’ This paper N/A
Recombinant DNA
pCAG4-Eco Addgene Addgene plasmid # 12260
pMX-GFP-Rac-G12V (Shi et al., 2018) Addgene Cat# 14567
ICOS cDNA ORF Clone Sino Biological MG50466-M
Software and Algorithms
FlowJo v9.3 FlowJo https://www.flowjo.com/
GraphPad Prism v6.0 GraphPad https://www.graphpad.com/scientific-software/prism/
Other
LSR Fortessa flow cytometer BD Biosciences N/A
MoFlow cell sorter Beckman-Coulter N/A
Reflection cell sorter iCyt N/A
ODYSSEY Fc Analyzer LI-COR N/A
Gene set enrichment analysis (GSEA) software suite (Subramanian et al., 2005) http://www.broadinstitute.org/gsea/msigdb/