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. 2020 May 19;52(5):794–807.e7. doi: 10.1016/j.immuni.2020.03.010
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Biotin B220, clone RA3-6B2 Biolegend Cat #: 103204;
RRID: AB_312989
Biotin Cd11b, clone M1/70 Biolegend Cat #: 101204
RRID: AB_312787
Biotin Ly6G, clone 1A8 Biolegend Cat #: 127604;
RRID: AB_1186108
Biotin CD49b, clone DX5 Biolegend Cat #: 108904;
RRID: AB_313411
PE CD45, clone 30F11 Biolegend Cat #: 103106;
RRID: AB_312971
PerCP/Cy5.5 TER119 Biolegend Cat #: 116227;
RRID: AB_893638
PE CD19, clone 6D5 Biolegend Cat #: 115508;
RRID: AB_313643
PE CD3, clone 17A2 Biolegend Cat #: 100206;
RRID: AB_312663
Alexa700 B220, clone RA3-682 Biolegend Cat #: 103232;
RRID: AB_493717
APC CD3, clone 145-2C11 Biolegend Cat #: 100236;
RRID: AB_2561456
BV785 CD4, clone RM4-5 Biolegend Cat #: 100552;
RRID: AB_2563053
BV605 CD8, clone 53-6.7 Biolegend Cat #: 100744;
RRID: AB_2562609
APC-Cy7 CD45, clone 30-F11 Biolegend Cat #: 103116;
RRID: AB_312981
APC-Cy7 TER119, clone TER-119 Biolegend Cat #: 116223;
RRID: AB_2137788
APC CD31, clone MEC13.3 Biolegend Cat #: 102410;
RRID: AB_312905
PE Podoplanin, GP38, clone 8.1.1 Biolegend Cat #: 127408;
RRID: AB_2161928
Integrin alpha4, CD49d, clone PS/2 BioXcell Cat #: BE0071;
RRID: AB_1107657
Integrin alphaL, CD11a, clone M17/4 BioXcell Cat #: BE0006;
RRID: AB_1107578
FITC CD169, clone 3D6.112 BIO-RAD Cat #: MCA884FT;
RRID: AB_1100895
GFP, rabbit polyclonal Thermo Fisher Cat #: A-11122;
RRID: AB_221569
Alkaline Phosphatase anti-Fluorescein, polyclonal Sigma Cat #: 11426338910 Roche;
RRID: AB_2734723
HRP anti-rabbit, polyclonal Thermo Fisher Cat #: 31460;
RRID: AB_228341
Alexa647 CD169, clone 3D6.112 BIO-RAD Cat #: MCA947A647;
RRID: AB_10545834
CD31, clone MEC13.3 BD Biosciences Cat #: 557355;
RRID: AB_396660
Biotin CD140b, PDGFR beta, clone APB5 Biolegend Cat #: 136010;
RRID: AB_2236916
ER-TR7, clone ER-TR7 BioXcell Cat #: BE0253;
RRID: AB_2687734
Biotin CCL21, polyclonal R&D systems Cat #: BAF457;
RRID: AB_2072082
Podoplanin, GP38, clone 8-1-1 eBioscience Cat #: 14-5381-82;
RRID: AB_1210505
Alexa488 anti-rat IgG Thermo Fisher Cat #: A21208;
RRID: AB_2535794
eFluor660 anti-rat IgG Thermo Fisher Cat #: 50-4017-82;
RRID: AB_2574211
Alexa488 anti-rabbit igG Thermo Fisher Cat #: A-11008;
RRID: AB_143165
Alexa647 anti-Armenian hamster IgG Jackson ImmunoResearch Cat #: 127-605-160;
RRID: AB-2339001
Biotin CD4 Biolegend Cat #: 100508;
RRID: AB-312711
Alexa647 B220, clone RA3-6B2 Biolegend Cat #: 103226;
RRID: AB-389330
Cy3 aplphaSMA, clone 1A4 Sigma Cat #: C6198-100UL;
RRID: AB-476856
Rabbit anti-EYFP living colors full length GFP, polyclonal Takara Cat #: 632592;
RRID: AB-2336883
Goat anti-CCL21 R&D systems Cat #: AF457;
RRID: AB-2072083
Rat anti-ER-TR7 Abcam Cat #: ab51824;
RRID: AB-881651
Alexa488 anti-rabbit IgG Jackson ImmunoResearch Cat #: 111-547-003;
RRID: AB-2338058
Dylight549 anti-rat IgG Jackson ImmunoResearch Cat #: 112-505-175;
RRID: AB-2338335
Dylight549 anti-Syrian Hamster IgG Jackson ImmunoResearch Cat #: 307-506-003;
RRID: AB-2339595
Critical Commercial Assays
Alexa488 streptavidin Invitrogen Cat #: S11223
Cy3 streptavidin Jackson ImmunoResearch Cat #: 016-160-084
Alexa647 streptavidin Jackson ImmunoResearch Cat #: 016-600-084
BioMag Goat anti-rat IgG QIAGEN Cat #: 310107
CFSE 5-(and 6-)-carboxyfluorescein diacetate succinimidyl ester Life Technologies Cat #: C1157
Cell tracker blue probe CMAC 7amino4chloromethylcoumarin Life Technologies Cat #: C2110
Cell Tracker orange CMTMR Life Technologies Cat #: C2927
Pertussis Toxin Islet Activating Protein Salt-Free (PTX) Quadratech Diagnostics Ltd. Cat #: 181
LPS, Escherichia coli 0111:B4 Sigma Aldrich Cat #: L4391
Dextran-FITC 2000KDa Sigma Aldrich Cat #: FD2000S
Collagenase P Sigma Aldrich Cat #: 11213857001
Dispase I Sigma Aldrich Cat #: D4818-2MG
MACS TER119 microbeads Miltenyi Biotec Cat #: 130-049-901
MACS CD45 microbeads Miltenyi Biotec Cat #: 130-052-301
Experimental Models: Organisms/Strains
Actin-CFP mice, Tg(ACTB-ECFP)1Nagy/J Jackson Laboratory 004218
Ccr7−/− mice, B6.129P2(C)-Ccr7tm1Rfor/J Jackson Laboratory 006621
Ubi-GFP mice, C57BL/6-Tg(UB C-GFP)30Scha/J Jackson Laboratory 004353
Rosa26-YFP mice, B6.129X1-Gt(ROSA)26Sortm1(EYFP)cos/J Jackson Laboratory 001648
CD19cre/cre mice, B6.129P2(C)-Cd19tm1(Cre)Cgn/J Jackson Laboratory 006785
Tlr4−/− mice, B6(Cg)-Tlr4tm1.2Karp/J Jackson Laboratory 029015
Ubi-PA-GFP mice, B6.Cg-PtprcaTg(UBC-PA-GFP)1Mnz/J Jackson Laboratory 022486
hCD2-DsRred mice Veiga-Fernandes et al., 2007 N/A
VE-cadherin-tdTomato, C57BL/6-Tg(Cdh5-cre/ERT2)rosa26-TdTomato Kindly provided by Dr. Bostjan MArkelc, Oxford University N/A
BAC-transgenic C57BL/6-Tg(Ccl19-tTA)687BIAT Available under MTA, laboratory of Dr. Burkhard Ludewig N/A
LC1 Schönig et al., 2002 N/A
Oligonucleotides
qPCR primers Ccl19 FP:
CTG CAA GAG AAC TGA ACA GAC		 Life Technologies N/A
qPCR primers Ccl19 RP:
CTT CTG ACT CTC TAG GTC TAC		 Life Technologies N/A
qPCR primers Ccl21 FP:
ATCGTGAAAGCCTTCGCTACCTT		 Life Technologies N/A
qPCR primers Ccl21 RP:
GCTGTTGCCTTTGTTCTTGGCAGA		 Life Technologies N/A
qPCR primers GAPDH FP:
AGGTCGGTGTGAACGGATTTG		 Life Technologies N/A
qPCR primers GAPDH RP:
TGTAGACCATGTAGTTGAGGTCA		 Life Technologies N/A
QuantiTect Primers Hprt QIAGEN QT00166768
QuantiTect Primers Ccl19 QIAGEN QT02532173
QuantiTect Primers Ccl21a QIAGEN QT00284753
Software and Algorithms
FlowJo 9.7.1 (Treestar inc) https://www.flowjo.com/ RRID: SCR_008520
Graphpad prism version 8 https://www.graphpad.com/scientific-software/prism/ RRID: SCR_002798
Adobe illustrator CS6 http://www.adobe.com/products/illustrator.html RRID: SCR_010279
Imaris 9.2.1 http://www.bitplane.com/imaris/imaris RRID: SCR_007370
Fiji, ImageJ, NIH Bethesda http://fiji.sc/ RRID: SCR_002285
ZEN Digital Imaging for Light Microscopy http://www.zeiss.com/microscopy/en_us/products/microscope-software/zen.html#introduction RRID: SCR_013672