Recombinant murine ccl19

5 μm pore size trans‐well plates (Corning Costar) were used. Upper chambers were loaded with 5 × 10⁴ BMDC, or FACS‐sorted CD11c⁺MHCII⁺ SI‐LP DC in 200 μL of IMDM supplemented with 10% FBS and 20 ng/mL recombinant murine GM–CSF (cIMDM), and the lower chambers with 500 μL of cIMDM with or without 50 ng/mL recombinant murine CCL19 (R&D Systems). To assess cell migration through 3D collagen, the upper chamber was loaded with 5 × 10⁴ BMDC or sorted CD11c⁺MHCII⁺ SI‐LP DC in cIMDM mixed 1:2 with Collagen type I (Millipore) supplemented with MEM 10X and NaHCO₃ to pH 7.2. Final concentration of Collagen type I was 3 mg/mL. Following incubation for 2 h, migrated cells were isolated from the bottom compartment and quantified by flow cytometry using Flow Count Fluorospheres (Beckman Coulter).

S1P was from Avanti Polar Lipids (Alabaster, AL). Recombinant murine CCL19, CCL5, CCL22, CCL2, CXCL12, CXCL10, IL-6, TNFα and IFNγ were from R&D Systems (Minneapolis, MN). Anti-VCAM-1 (clone 429), anti-ICAM-1 (clone YN1/1), anti-CD4 (GK1.5), anti-CD25 (clone PC61.5), anti-CD44 (clone IM7) and anti-TNFα were from eBioscience (San Diego, CA). Anti-human CD31, anti-human LYVE1, anti-human VEGFR3, anti-human GP38, anti-human ICAM-1 and anti-human VCAM-1 antibodies were from Biolegend (San Diego, CA). Anti-VE-cadherin was from BD PharMingen (San Diego, CA). AlexFluor-555 phalloidin was from Life Technologies (Grand Island, NY). Anti-moesin was from Cellsignaling (Danvers, MA). Anti-β-catenin, anti-collagen and anti-laminin were from Abcam (Cambridge, UK). Anti-prox1 antibody was from Bioss (Woburn, MA). Recombinant human laminin 511 was from Biolamina AB (Sundbyberg, Sweden). EIA grade gelatin was from Bio-Rad (Berkely, CA).