Fig. 4. Thymic CD301b⁺ cDC2 require type 2 cytokines.

(a) Heatmap displaying the relative expression of leading-edge genes from the PID_IL4_2PATHWAY gene set among sorted thymic XCR1⁺ cDC1, CD301b⁻ cDC2, and CD301b⁺ cDC2 from 8-week-old C57BL/6 mice. (b) Gene set enrichment analysis showing the enrichment for IL-4 signature genes among sorted thymic CD301b⁻ cDC2 and CD301b⁺ cDC2. NES, normalized enrichment score. FDR q-value, false discovery rate. (c) Heatmap displaying the relative expression of leading-edge genes from the GNF2_STAT6 gene set among sorted thymic XCR1⁺ cDC1, CD301b⁻ cDC2, and CD301b⁺ cDC2 from 8-week-old C57BL/6 mice. (d) Gene set enrichment analysis showing the enrichment of STAT6 related genes among sorted thymic CD301b⁻ and CD301b⁺ cDC2. NES, normalized enrichment score. FDR q-value, false discovery rate. (e) Representative flow cytometry of thymic XCR1⁺ cDC1, CD301b⁻ cDC2, and CD301b⁺ cDC2 stained for PDL2 (left) and PDL2 gMFI (right) in C57BL/6 mice (n=3). (f) Representative flow cytometry of XCR1⁺ cDC1 (top; magenta gate) and SIRPα⁺ cDC2 (top; teal gate) and CD301b⁺ DC2 (bottom; green gate) from BALB/c.BYJ (left), IL-4 KO (middle left), IL-13 KO (middle; from separate experiment), IL-4Rα KO (middle right), and CD1d KO (right) mice, all on a BALB/c background. Numbers adjacent to outlined areas represent percent cells in each. (g) Frequency of CD301b⁺ among SIRPα⁺ cDC2 in thymus of BALB/c.BYJ (n=15), IL-4 KO (n=10), IL-13 KO (n=4), IL-4Rα KO (n=8), and CD1d KO (n=6) mice. Six to twelve-week-old male and female mice (e, f) were used. Each dot represents an individual mouse (e, g). Small horizontal lines indicate the mean, and error bars represent SD. Data are representative of at least three independent experiments (e, f), are pooled from at least three independent experiments (g). ns=not significant, *P<0.05, ****P< 0.0001. One-way ANOVA with Tukey’s multiple comparisons test was used.