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 |