REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Armenian Hamster monoclonal anti-CD3ε (clone 145-2C11) APC conjugated	BD Biosciences	Cat# 553066, RRID:AB_398529
Armenian Hamster monoclonal anti-CD3ε (clone 145-2C11) PE conjugated	BD Biosciences	Cat# 553063, RRID:AB_394596
Goat polyclonal anti-CD3ε	Santa Cruz Biotechnology	Cat# sc-1127, RRID:AB_631128
Rat monoclonal anti-CD4 (clone RM4-4)	BD Biosciences BioLegend	Cat#: 553053, RRID: AB_394588
		Cat#: 116018, RRID: AB_2650936
Rat monoclonal anti-CD4 (clone RM4-4) FITC conjugated	BioLegend	Cat#: 116003, RRID: AB_313688
Rat monoclonal anti-CD4 (clone RM4-4) PE conjugated	BioLegend	Cat# 116006, RRID: AB_313691
Rat monoclonal anti-CD4 (clone RM4-5) biotin conjugated	BD Biosciences	Cat# 553649, RRID:AB_394969
Rat monoclonal anti-CD4 (clone RM4-5) PE conjugated	BD Biosciences	Cat# 553049, RRID:AB_394585
Rat monoclonal anti-CD4 (clone RM4-5) BV650 conjugated	BioLegend	Cat# 100545, RRID:AB_11126142
Rat monoclonal anti-CD4 (clone RM4-5) APC conjugated	BD Biosciences	Cat# 553051, RRID:AB_398528
Rat monoclonal anti-CD4 (clone RM4-5) AF700 conjugated	BD Biosciences	Cat# 557956, RRID:AB_396956
Rat monoclonal anti-CD4 (clone H129.19) PE conjugated	BD Biosciences	Cat# 553652, RRID:AB_394972
Rat monoclonal anti-CD4 (clone H129.19) FITC conjugated	BD Biosciences	Cat# 553651, RRID:AB_394971
Rat monoclonal anti-CD4 (clone GK1.5) Alexa Fluor 488 cojungated	Biolegend	Cat# 100423, RRID:AB_389302
Rabbit monoclonal anti-CD4 (clone D7D2Z)	Cell Signaling Technology	Cat# 25229, RRID:AB_2798898
Rat monoclonal anti-CD4 (clone YTS 177.9) biotin conjugated	Tomas Brdicka’s lab	N/A
Rat monoclonal anti-CD8a (clone 53-5.7) FITC conjugated	BD Biosciences BioLegend	Cat# 553032, RRID:AB_394570
		Cat# 100706, RRID:AB_312745
Rat monoclonal anti-CD8a (clone 53-5.7) BV421 conjugated	BioLegend	Cat# 100737, RRID:AB_10897101
Rat monoclonal anti-CD8a (clone 53-5.7) PE conjugated	BD Biosciences	Cat# 553033, RRID:AB_394571
Rabbit monoclonal anti-CD8a (clone D4W2Z)	Cell Signaling Technology	Cat# 98941, RRID:AB_2756376
Rat monoclonal anti-CD8b.2 (clone 53-5.8)	BD Biosciences	Cat# 553038, RRID:AB_394574
Rat monoclonal anti-CD8b.2 (clone 53-5.8) Biotin cojugated	BD Biosciences	Cat# 553039, RRID:AB_394575
Rat monoclonal anti-CD8b.2 (clone 53-5.8) FITC conjugated	BD Biosciences	Cat# 553040, RRID:AB_394576
Rat monoclonal anti-CD8b.2 (clone 53-5.8) PerCP-Cy5.5 conjugated	BioLegend	Cat# 140417, RRID:AB_2800650
Rat monoclonal anti-CD8b (clone eBioH35-17.2) PE-Cy7 conjugated	Thermo Fisher Scientific	Cat# 25-0083-82, RRID:AB_11218494
Rat monoclonal anti-CD11b (clone YBM 15.1.6) biotin conjugated	Tomas Brdicka’s lab	N/A
Rat monoclonal anti-CD25 (clone PC61) PE-Cy7 conjugated	BioLegend	Cat# 102016, RRID:AB_312865
Rat monoclonal anti-CD44 (clone IM7) PerCP-Cy5.5 conjugated	BioLegend	Cat# 103032, RRID:AB_2076204
Rat monoclonal anti-CD44 (clone IM7) BV650 conjugated	BioLegend	Cat# 103049, RRID:AB_2562600
Rat monoclonal anti-CD44 (clone IM7) biotin conjugated	BioLegend	Cat# 103003, RRID:AB_312954
Mouse monoclonal anti-CD45.2 (clone 104) APC-Cy7 conjugated	BD Biosciences	Cat# 560694, RRID:AB_1727492
Mouse monoclonal anti-CD45.2 (clone 104) Alexa Fluor 700 conjugated	Biolegend	Cat# 109822, RRID:AB_493731
Rat monoclonal anti-CD45R/B220 (clone RA3-6B2) biotin conjugated	BD Biosciences	Cat# 553085, RRID:AB_394615
Rat monoclonal anti-CD62L (clone MEL-14) PE-Cy7 conjugated	BioLegend	Cat# 104418, RRID:AB_313103
Armenian Hamster monoclonal anti-CD69 FITC conjugated	BD Biosciences	Cat# 553236, RRID:AB_394725
Mouse monoclonal anti-Lck (clone 3a5)	Santa Cruz Biotechnology	Cat# sc-433, RRID:AB_627880
Mouse monoclonal anti-Lck (clone 3a5) PE conjugated	Santa Cruz Biotechnology	Cat# sc-433 PE, RRID: N/A
Mouse monoclonal anti-MHCI (H2Kb; clone Y3.8)	Ed Palmer’s lab, University of Basel	N/A
Armenian Hamster monoclonal anti-TCRβ (clone H57-597) APC conjugated	BD Biosciences Biolegend	Cat# 553174, RRID:AB_398534
		Cat# 109212, RRID:AB_313435
Rabbit monoclonal anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (clone D13.14.4E)	Cell Signaling Technology	Cat# 4370, RRID:AB_2315112
Mouse monoclonal anti-Src, non-phospho (Tyr416) (clone 7G9)	Cell Signaling Technology	Cat# 2102, RRID:AB_331358
Rabbit polyclonal anti-Phospho-Src Family (Tyr416)	Cell Signaling Technology	Cat# 2101, RRID:AB_331697
Mouse monoclonal anti-Phosphotyrosine (clone 4G10) PE conjugated	Millipore	Cat# FCMAB323PE, RRID:AB_10805942
Mouse monoclonal anti-CD247 phospho (Tyr142) (clone K25-407.69) PE conjugated	BD Biosciences	Cat# 558448, RRID:AB_647237
Rabbit monoclonal anti-Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (clone 65E4)	Cell Signaling Technology	Cat# 2717, RRID:AB_2218658
Mouse monoclonal anti-Lamin B1 (clone 119D5-F1)	Santa Cruz Biotechnology	Cat# sc-56143, RRID:AB_2136302
Mouse monoclonal anti-Zap70 (clone 1E7.2) Alexa Fluor 488 conjugated	Thermo Fisher Scientific	Cat# MHZAP7020, RRID:AB_10375316
Rabbit polyclonal anti-GAPDH	Sigma-Aldrich	Cat# G9545, RRID:AB_796208
Donkey polyclonal anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 conjugated	Thermo Fisher Scientific	Cat# A-31572, RRID:AB_162543
Goat polyclonal anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 conjugated	Thermo Fisher Scientific	Cat# A-21429, RRID:AB_2535850
Goat polyclonal anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 conjugated	Thermo Fisher Scientific	Cat# A-11034, RRID:AB_2576217
Goat polyclonal anti-mouse IgG (H+L) Secondary Antibody, HRP conjugated	Jackson ImmunoResearch Labs	Cat# 115-035-003, RRID:AB_10015289
Donkey polyclonal anti-Goat IgG (H+L) antibody, HRP conjugated	Jackson ImmunoResearch Labs	Cat# 705-035-003, RRID:AB_2340390
Goat polyclonal anti-rabbit IgG (H+L) Secondary Antibody, HRP conjugated	Jackson ImmunoResearch Labs	Cat# 111-035-003, RRID:AB_2313567
Bacterial and Virus Strains
Listeria monocytogenes (transfected with empty pPL1,pPL2)	(Zehn et al., 2009)	N/A
Listeria monocytogenes expressing SIINFEKL (OVA) peptide (transfected with pPL1,pPL2 carrying OVA peptide coding sequence)	(Zehn et al., 2009)	N/A
Listeria monocytogenes expressing SIITFEKL (T4) peptide (transfected with pPL1,pPL2 carrying T4 peptide coding sequence)	(Zehn et al., 2009)	N/A
Listeria monocytogenes expressing SIIQFEHL (Q4H7) peptide (transfected with pPL1,pPL2 carrying Q4H7 peptide coding sequence)	(King et al., 2012)	N/A
Listeria monocytogenes expressing FEAQKAKANKAVD (3K) peptide (transfected with pPL1,pPL2 carrying 3K peptide coding sequence)	This paper	N/A
Listeria monocytogenes expressing FEAAKAKANKAVD (P2A) peptide (transfected with pPL1,pPL2 carrying P2A peptide coding sequence)	This paper	N/A
Listeria monocytogenes expressing FAAQKAKANKAVD (P-1A) peptide (transfected with pPL1,pPL2 carrying P-1A peptide coding sequence)	This paper	N/A
Chemicals, Peptides, and Recombinant Proteins
AccuCheck Counting Beads	Thermo Fisher Scientific	Cat# PCB100
Qdot 605 Streptavidin Conjugate	Invitrogen	Cat# Q10103MP
CML Latex Beads, 4% w/v, 5 μm	Thermo Fisher Scientific	Cat# C37255
Nonidet P 40 Substitute	Sigma Aldrich	Cat# 74385
n-Dodecyl-beta-Maltoside Detergent	Thermo Fisher Scientific	Cat# 89903
cOmplete, EDTA-free Protease Inhibitor Cocktail Tablets	Roche	Cat# 05056489001
4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride	Sigma Aldrich	Cat# A8456
PhosSTOP	Roche Molecular Systems, Inc	Cat# 04906837001
Amersham Protran 0.45 NC nitrocellulose western blotting membranes	GE Healthcare	Cat# 10600002
Streptavidin Mag Sepharose	GE Healthcare	Cat# 28985738
LPS E.Coli O111:B4	Sigma Aldrich	Cat# LPS25
OVA peptide (SIINFEKL)	Eurogentec	Ref# AS-60193-1
T4 peptide (SIITFEKL)	Eurogentec	Ref# AS-64403
Q4H7 peptide (SIIQFEHL)	Eurogentec	Ref# AS-64405
3K peptide (FEAQKAKANKAVD)	Peptides and Elephants	N/A
P2A peptide (FEAAKAKANKAVD)	Peptides and Elephants	N/A
P-1A peptide (FAAQKAKANKAVD)	Peptides and Elephants	N/A
(+)-Biotin N-hydroxysuccinimide ester	Sigma Aldrich	Cat# H1759
Sephadex® G-25	Sigma Aldrich	Cat# S5772
Critical Commercial Assays
LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit	Thermo Fisher Scientific	L34976
CellTrace CFSE Cell Proliferation Kit	Thermo Fisher Scientific	C34554
Untouched Mouse CD8 Cells Kit	Dynabeads	Cat# 11417D
Untouched Mouse CD4 Cells Kit	Dynabeads	Cat# 11415D
Biotin Binder	Dynabeads	Cat# 11047
EasySep Mouse CD8 T Cell Enrichment Kit	Stem Cell	Cat# 19753A
RNA Clean and Concentrator-5	Zymo Research	Cat# R1013
Neuraminidase from Vibrio cholera, type II	Sigma Aldrich	Cat# N6514
Experimental Models: Cell Lines
Lutz	N/A	N/A
Experimental Models: Organisms/Strains
Mouse: C57BL/6J	Animal Facility of Institute of Molecular Genetics	JAX 000664
Mouse: C57BL/6J CD45.1	(Shen et al., 1985)	JAX 002014
Mouse: CD3ε−/−	(Sommers et al., 2000)	JAX 004177
Mouse: CD8.4	(Erman et al., 2006)	N/A
Mouse: OT-I Rag2−/−	(Hogquist et al., 1994, Shinkai et al., 1992)	N/A
Mouse: B3K508 Rag2−/−	(Huseby et al., 2005, Shinkai et al., 1992)	N/A
Mouse: FoxP3−/−	(Lin et al., 2005)	JAX 019933
Mouse: Nur77-GFP	(Moran et al., 2011)	JAX 016617
Mouse: FoxP3-GFP	(Fontenot et al., 2005)	N/A
Mouse: FoxP3-DTR	(Kim et al., 2007)	JAX 016958
Mouse: Lck−/−	This paper	N/A
Oligonucleotides
CD8 Forward – CCGTGGCTCAGTGAAGGGG	Sigma Aldrich	N/A
CD8’ Reverse – CTGACTAGCGGCTGTGGTAGC	Sigma Aldrich	N/A
CD8 Full length Reverse – CATTTGCAAACACGCTTTCGGCTC	Sigma Aldrich	N/A
CD8 Total Reverse - CTTGCCTTCCTGTCTGACTAGC	Sigma Aldrich	N/A
gRNA for Lck−/− generation: TTGCTGTCCAGTGGGACTAT GGG	N/A	N/A
Software and Algorithms
Source code for ‘Lck come&stay/signal duration’ model (MATLAB)	This paper (Data S1)	N/A
GraphPad Prism 5.04	GraphPad Software	N/A
FlowJo V9 and V10	FlowJo, LCC	N/A
R Studio V1.2.1335	RStudio, Inc.	N/A
Tescan Q-Phase software V7.727	Tescan Orsay Holding, a.s	N/A
Fiji (ImageJ version 1.52i)	Open Source	N/A
MATLAB	MathWorks	N/A
Other
LSRII	BD Biosciences	N/A
CantoII	BD Biosciences	N/A
FACSymphony	BD Biosciences	N/A
LSRFortessa	BD Biosciences	N/A
Influx Sorter	BD Biosciences	N/A
Cytek™ Aurora	Cytek	N/A
LI-COR Odyssey infrared imaging system	LI-COR Biosciences	N/A
Azure c200 imaging system	Azure Biosystems	N/A
Z2 Coulter Counter Analyzer	Beckman Coulter	N/A
LightCycler® 480 Instrument II	Roche Molecular Systems, Inc	N/A