Figure 4. CX₃CR1⁺Tstem-l have a distinct transcriptional signature.

(A-B) Ly108, CX₃CR1 and CD101 were used to distinguish and sort four T cell subsets from WT mice for analysis of differentially expressed genes using NanoString. (A) Number of differentially expressed genes between CX₃CR1⁺Tstem-l and other T cell subsets. (B) Heatmap representing relative expression of selected differentially expressed genes between all subsets. (C-F) Congenically marked CX₃CR1⁻Ly108⁺CD44⁺PD1⁺ CD8⁺ T cells (Tstem-l) were sorted from d17 Cl13-infected mice, labelled with cell tracker violet (CTV) and transferred into infection matched recipients. (C) Experimental scheme. (D) Proliferation and differentiation of Tstem-l in the spleen on d8 post-transfer. (E) Frequency of distinct subsets among undivided and divided cells. (F) Representative flow cytometry plot showing CX₃CR1 and CTV expression on transferred cells that remained TCF1 positive. Data are from at least two independent experiments (n=3–4 per time point). Data in B were analyzed with the use of nSolver and multiple comparison test with Benjamini-Yekutieli False Discovery Rate method; *, P < 0.05; **, P < 0.01; ***, P < 0.001. See also Figure S2.