Fig. 1: Runx3WRPW inhibits the development of CD8 T cells.
a, Dot plots showing CD4 and CD8α expression in mature thymocytes and spleen T cells (left) and quantification of percentage and number of CD8 SP thymocytes and splenocytes (right) in Runx3 (Rx3)+/+ ((thymus, n = 9; spleen, n = 10) Rx3+/W (thymus, n = 9; spleen, n = 10) and Rx3W/W (thymus, n = 9; spleen, n = 6) mice. Data are presented as mean ± standard deviation (s.d.). **P < 0.01, ***P < 0.0001, ****P < 0.0001 (one-way ANOVA with Tukey’s multiple comparison). b, Histogram (left) and quantification (right) of IL7Rα expression on CD8 SP thymocytes from Rx3+/+ (n = 5, Rx3+/W (n = 4) and Rx3W/W (n = 3) mice. Data represent mean ± s.d. *P < 0.05, **P < 0.01 (one-way ANOVA with Tukey’s multiple comparison). c, Pie charts showing the frequency of cells in the G0/G1, S and G2/M cell cycle stage in CD8 SP thymocytes from Rx3+/+(n = 3), Rx3+/W (n = 3) and Rx3W/W (n = 3) mice. Data represent mean ± s.d. *P < 0.0001 compared to Rx3+/+ (one-way ANOVA with Tukey’s multiple comparison). d, Heatmap showing the expressions of differentially expressed genes (fold-change > 2, FDR < 0.01) between CD8 SP thymocytes from Rx3+/+ (n = 2) and Rx3+/W (n = 3) mice assessed by bulk RNA-sequencing analysis (left) and graph showing statistically significant enrichment of GO biological process annotations among downregulated genes in Rx3+/W mature CD8 SP thymocytes (Fold-change ≥ 2, FDR ≤ 0.01). e, Normalized count of Tnk2, Il7r, Cd8a and CD8b1 (left) and expression of Tnk2 mRNA relative to Hprt mRNA (right) in CD8 SP thymocytes from Rx3+/+ (n = 2 and Rx3+/W (n = 3) mice. Data represent mean ± s.d. ****P < 0.0001 (unpaired Student’s t-test, two-sided). f, ChIP-seq tracks showing bindings of histone H3K27me3, H3K4me3 and Runx3 at the Tnk2 locus in CD8 SP thymocytes from Rx3+/+ (n = 1) and Rx3+/W (n = 1) mice. A single experiment was performed. g, Venn diagrams showing the H3K27me3 regions in the whole genome between CD8 SP thymocytes from Rx3+/+ (n = 1 and Rx3+/W (n = 1) mice (left) and histogram showing H3K27me3 signals on regions detected specifically in Rx3+/W CD8 SP thymocytes.
