Figure 1. CCR7-dependent entry of ILCs into peripheral LNs.

To assess the migratory kinetics of different cellular populations within peripheral LNs, the bLN of Kaede mice was photoconverted and analysed 72 hrs later. (A) Expression of Kaede Green versus Kaede Red protein by CD45⁺ cells isolated from the bLN at 72 hrs post photoconversion. (B) Percentage of CD45⁺ cells expressing Kaede Green or Kaede Red protein at 72 hrs post photoconversion (n=10). (C) Characterisation of Kaede Green⁺ CD45⁺ cells isolated from the bLN, showing αβ T cells (CD45⁺ CD3⁺ TCRβ⁺ TCRγδ^-), γδ T cells (CD45⁺ CD3⁺ TCRβ^- TCRγδ⁺), ILCs (CD45⁺ IL-7Rα⁺ Lin{B220, CD11b, CD11c, CD3, CD5, CD19, Ter119, Gr1, CD49b, F4/80, FcεRI}^-) and LCs (CD45⁺ MHCII⁺ CD11c⁺ CD207⁺). (D) Percentage of αβ T cells (n=7), γδ T cells (n=7), ILCs (n=10) or LCs (n=5) amongst CD45⁺ Kaede Green⁺ cells. (E) Characterisation of Kaede Red⁺ cells isolated from the bLN, showing αβ T cells, γδ T cells, ILCs and LCs. (F) Percentage of αβ T cells (n=7), γδ T cells (n=7), ILCs (n=10) and LCs (n=5) amongst CD45⁺ Kaede Red⁺ cells. Expression of Kaede Green versus Kaede Red protein by αβ T cells, γδ T cells, ILCs and LCs isolated from the bLN of (G) Kaede and (H) CCR7^-/- Kaede mice. (I) Number of αβ T cells (WT n=7, CCR7^-/- n=8), γδ T cells (WT n=7, CCR7^-/- n=8), ILCs (WT n=6, CCR7^-/- n=8) and LCs (WT=5, CCR7^-/- n=7) isolated from the bLN of WT Kaede versus CCR7^-/- Kaede mice. (J) Number of Kaede Green αβ T cells (WT n=7, CCR7^-/- n=8), γδ T cells (WT n=7, CCR7^-/- n=8), ILCs (WT n=7, CCR7^-/- n=8) and LCs (WT n=5, CCR7^-/- n=7) isolated from the bLN. Data are pooled from a minimum of 2 independent experiments. Values on flow cytometry plots represent percentages, bars on scatter plots represents the median, which is also shown numerically. Statistical significance was tested using an unpaired, non-parametric, Mann-Whitney two-tailed U test: *p≤0.05, **p≤0.01, ***p≤0.001.