CX₃CR1-GFP⁺ cells upregulate PD-L2 as they extravasate and accumulate in liver tissue.

<p>(<b>A and B</b>) Representative contour plots and graphs displaying PD-L2 expression on liver leukocytes isolated from uninfected or infected <i>CX₃CR1^gfp/+</i> mice gated on live, single, lineage (CD3, B220, DX5 and Siglec F) negative, SSC^low cells. n = 4 mice/group. (<b>C</b>) qRT-PCR analysis of Relmα expression. (<b>D</b>) Representative contour plots show blood/tissue partitioning of GFP⁺ liver leukocytes from CX₃CR1^GFP/+ mice using <i>in vivo</i> CD45 staining. Plots display cells gated on single, live, lineage negative cells. (<b>E</b>) The proportion of CD45⁺ (white) or CD45⁻ (black) cells gated on live single, lineage⁻, GFP⁺ cells. (<b>F</b>) Contour plots display blood/tissue partitioning of PDL2⁺, GFP⁺ cells gated on single, live, lin-, CX₃CR1-GFP⁺ cells. (<b>G</b>) Proportions of GFP⁺PDL2⁺ cells that are CD45⁺ (white) or CD45⁻ (black) as in (<b>E</b>). (<b>H</b>). qRT-PCR analysis of YM1 expression in liver leukocytes sorted from 8 week infected mice using <i>in vivo</i> CD45 staining to isolate CD45−(tissue) GFP+ granuloma macrophages from CD45+ (blood) GFP+, Ly6C^high and Ly6C^low monocytes. CD11b-GFP− cells from infected mice were used as a negative control. Dots represent individual mice from one of two experiments. Statistical significance was determined by One-way ANOVA with Dunnett's test. ** = p<0.001, *** = p<0.0001. Results are representative of at least two experiments that include n = 3–4 mice/group.