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) | Thermo Fisher Scientific | Cat# 25-0621-81; 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-CD122 (TM-β1) | Thermo Fisher Scientific | Cat# 12-1222-81; RRID:AB_465835 |
| Anti-CD25 (PC61.5) | Thermo Fisher Scientific | Cat# 17-0251-82; RRID:AB_469366 |
| Anti-CXCR5 (2G8) | BD Bioscience | Cat# 551960; RRID:AB_394301 |
| Anti-Streptavidin-PE | BioLegend | Cat# 410504; RRID:AB_2571915 |
| 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-pY694-STAT5 (Flow cytometry) | BD Bioscience | Cat# 612567; RRID:AB_399858 |
| 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-Caspase 3-PE | BD Bioscience | Cat# 557091; RRID:AB_396582 |
| 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-IL-2 (JES6-1) | Bio-X-Cell | Cat# BE0043; RRID:AB_1107702 |
| Anti-pY694-STAT5 (Immunoblot) | Cell Signaling Technology | Cat# 9351; RRID:AB_2315225 |
| Anti-pY694-STAT5 (D47E7) (Immunofluorescence) | Cell Signaling Technology | Cat# 4322P; RRID:AB_10548756 |
| Anti-Lamin B1 (D9V6H) | Cell Signaling Technology | Cat# 13435; RRID:AB_2737428 |
| Anti-β-actin (AC-74) | Sigma-Aldrich | Cat# A2228; RRID:AB_476697 |
| Anti-STAT5 | Cell Signaling Technology | Cat# 9363; RRID:AB_2196923 |
| Anti-phospho-S6 (S235/236) (D57.2.2E) | Cell Signaling Technology | Cat# 4858; RRID:AB_916156 |
| Anti-phospho-ERK1/2 (T202/204) (D13.14.4E) | Cell Signaling Technology | Cat# 4370; RRID:AB_2315112 |
| Anti-phospho-Akt (S473) | Cell Signaling Technology | Cat# 4060 |
| Anti-phospho-Mob1 (T35) (D2F10) | Cell Signaling Technology | Cat# 8699S; RRID:AB_11139998 |
| Anti-phospho-Mst1/2 (T183/180) | Cell Signaling Technology | Cat# 3681S; RRID:AB_330269 |
| Anti-Mst1 (immunoblot) | Cell Signaling Technology | Cat# 3682; RRID:AB_2144632 |
| Anti-Mst1 (EP1465Y, Immunoprecipitation) | Abcam | Cat# ab51134; RRID:AB_881247 |
| Anti-DOCK8 (H-159) | Santa Cruz | Cat# sc-292124; RRID:AB_10845110 |
| Anti-GFP-Booster_ATTO488 | ChromoTek | Cat# gba488-100; RRID:AB_2631386 |
| Anti-B220-Alexa Fluor 594 (RA3-6B2) | BioLegend | Cat# 103254; RRID:AB_2563229 |
| Anti-CD3-Alexa Fluor 647 (17A2) | BioLegend | Cat# 100209; RRID:AB_389323 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| 7-AAD | Sigma-Aldrich | Cat# A9400 |
| EHT-1864 (Rac inhibitor) | Cayman Chemical | Cat# 17258; CAS: 754240-09-0 |
| EHop-016 (Rac inhibitor) | Tocris | Cat# 6248; CAS: 1380432-32-5 |
| Cytochalasin D (actin inhibitor) | Tocris | Cat# 1233; CAS: 22144-77-0 |
| Colchicine (microtubule inhibitor) | Tocris | Cat# 1364; CAS: 64-86-8 |
| Tofacitinib (JAK3 inhibitor) | Tocris | Cat# 4556; CAS: 540737-29-9 |
| PS-1145 (IKKβ inhibitor) | Cayman Chemical | Cat# 14862; CAS: 431898-65-6 |
| XMU-MP-1 (Mst1 inhibitor) | Selleck Chemicals | Cat# S8334; Lot# 01; CAS: 2061980-01-4 |
| U-0126 (ERK inhibitor) | Cayman Chemical | Cat# 70970; CAS: 109511-58-2 |
| SB203580 (p38 MAPK inhibitor) | EMD Millipore | Cat# 559389; CAS: 152121-47-6 |
| GSK2334470 (PDK1 inhibitor) | Sigma-Aldrich | Cat# SML0217; CAS: 1227911-45-6 |
| Akt inhibitor VIII (Akt inhibitor) | EMD Millipore | Cat# 124018; CAS: 612847-09-3 |
| Rapamycin (mTORc1 inhibitor) | EMD Millipore | Cat# 553210; CAS: 53123-88-9 |
| SP600125 (JNK inhibitor) | Sigma-Aldrich | Cat# S5567; CAS: 129-56-6 |
| LY294002 (PI3K inhibitor) | Sigma-Aldrich | Cat# 440202; CAS: 154447-36-6 |
| SB216763 (GSK3 inhibitor) | Tocris | Cat# 1616; CAS: 280744-09-4 |
| PF 3644022 (MK2 inhibitor) | Tocris | Cat# 4279; CAS: 1276121-88-0 |
| MNK1 inhibitor | EMD Millipore | Cat# 454861; CAS: 522629-08-9 |
| SB 747651A (MSK1 inhibitor) | Tocris | Cat# 4630; CAS: 1781882-72-1 |
| Phalloidin-Alexa Fluor 647 | Thermo Fisher Scientific | Cat# A2287 |
| Recombinant murine IL-2 (complex with anti-IL-2) | Peprotech | Cat# 212-12 |
| Recombinant human IL-2 (in vitro stimulation) | R&D Systems | Cat# 202-IL |
| Phorbol 12-myristate 13-acetate (PMA) | Sigma-Aldrich | Cat# P8139 |
| Ionomycin | Sigma-Aldrich | Cat# I0634 |
| Golgi-STOP | BD Bioscience | Cat# 554724 |
| Recombinant human IL-6 | National Cancer Institute | Cat# NSC720836 |
| Recombinant murine IL-3 | Thermo Fisher Scientific | Cat# PMC0034 |
| Recombinant murine SCF | Thermo Fisher Scientific | Cat# PMC2115 |
| Critical Commercial Assays | ||
| Rac1 Pull-down Activation Assay Biochem Kit | Cytoskeleton | Cat# BK035 |
| Mouse Anti-dsDNA Ig's (Total A+G+M) ELISA Kit, 96 tests, Quantitative | Alpha Diagnostic International | Cat# 5110 |
| Foxp3 transcription Factor Staining buffer set | Thermo Fisher Scientific | Cat# 00-5523-00 |
| Cytofix/Cytoperm Fixation/Permeabilization Solution Kit | BD Bioscience | Cat# 554714 |
| Annexin V Apoptosis Detection Kit | BD Bioscience | Cat# 559763 |
| Pierce BCA Protein Assay Kit | Thermo Fisher Scientific | Cat# 23225 |
| Lineage cell depletion kit | Miltenyi Biotec | Cat# 130-090-858 |
| Deposited Data | ||
| Treg Microarray data | This paper | GEO: GSE120711 |
| Experimental Models: Organisms/Strains | ||
| Mouse: C57BL/6J | The Jackson Laboratory | Cat# 000664; RRID:IMSR_JAX:000664 |
| Mouse: Stk4f/fStk3f/f: Stk4tm1.1Rjo Stk3tm1.1Rjo/J | The Jackson Laboratory | Cat# 017635; RRID:IMSR_JAX:017635 |
| Mouse: Yap1f/f | E. Olson (Xin et al., 2013) | N/A |
| Mouse: Wwtr1f/f | E. Olson(Xin et al., 2013) | N/A |
| Mouse: CD45.1+: B6.SJL-Ptprca Pep3b/BoyJ | The Jackson Laboratory | Cat# 002014; RRID:IMSR_JAX:002014 |
| Mouse: ROSA26GFP: Gt(ROSA)26Sortm2Sho/J | The Jackson Laboratory | Cat# 004077; RRID:IMSR_JAX:004077 |
| Mouse: Cd4Cre: Tg(Cd4-cre)1Cwi/BfluJ | The Jackson Laboratory | Cat# 017336; RRID:IMSR_JAX:017336 |
| Mouse: Tnfrsf4Cre: Tnfrsf4tm2(cre)Nik/J | The Jackson Laboratory | Cat# 012839; RRID:IMSR_JAX:012839 |
| Mouse: Ifng−/−: Ifngtm1Ts/J | The Jackson Laboratory | Cat# 002287; RRID:IMSR_JAX:002287 |
| Mouse: Rag1−/−: Rag1tm1Mom/J | The Jackson Laboratory | Cat# 002216; RRID:IMSR_JAX:002216 |
| Mouse: Foxp3YFP-Cre | A. Rudensky (Chinen et al., 2016) | N/A |
| Recombinant DNA | ||
| Mouse STAT5bCA | Y. Feng (Feng et al., 2014) | N/A |
| pmCherry-STAT5CA | This paper | N/A |
| pMX-GFP-Rac-G12V | (Hall et al., 2006) | Addgene Cat# 14567 |
| pMSCV-IRES-Thy1.1-Rac1N17 | This paper | N/A |
| Software and Algorithms | ||
| FlowJo V9.3 | FlowJo | https://www.flowjo.com/ |
| Prism6 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Image J | NIH | https://imagej.nih.gov/ij/ |
| Other | ||
| LSR II flow cytometer | BD Bioscience | N/A |
| LSR Fortessa flow cytometer | BD Bioscience | N/A |
| MoFlow | Beckman-Coulter | N/A |
| Reflection | i-Cyt | N/A |