Fig. 5

PD-1 is a negative regulator of Nr4a1⁺ILC2s. a The dot plot shows the expression of immune checkpoint molecules in Nr4a1⁺ILC2s. b Left panel: t-SNE plot showing the clustering of 703 ILC2s (362 KO and 341 WT). Right panel: t-SNE plot showing the time points information about both WT and KO ILC2s. c Experimental design for the examination of Nr4a1⁺ILC2s after PD-1 KO. d FACS analysis of lung ILC2s of Nr4a1^eGFPcre/+ mice and Pdcd1^−/−;Nr4a1^eGFPcre/+ mice on d7 after papain challenge. e The percentages of Nr4a1⁺ILC2s in Nr4a1^eGFPcre/+mice (n = 3) and Pdcd1^−/−;Nr4a1^eGFPcre/+ mice (n = 3). Data are presented as means ± SEM. The significance between means was analyzed by unpaired t-test. f The box plot shows distributions of memory scores, Trm scores, Tcm/Tem scores, and wounding healing scores of clusters from both WT and PD-1 KO ILC2s. g The bar chart shows the GO pathways analysis of DEGs (P value < 0.05 and log2FC > 1) between PD-1 KO Cm4 and WT Cm4 ILC2s. h The percentage of Il5-expressing ILC2s in Nr4a1⁺ILC2s from Pdcd1^−/−;Il5^tdtomato/+ mice or Il5.^tdtomato/+mice. i Proportions of Cm1 and Cm2 cells in WT non-activation cells or in PD-1 KO non-activation cells. j Proportions of Cm3 and Cm4 cells in WT activation cells or in PD-1 KO activation cells. k The box plot shows the distribution of proliferation scores of clusters in WT and PD-1 KO ILC2s. l The top 2 panels show the WT ILC2 activation trajectory, colored by cluster (left) and inferred by pseudotime (right). The bottom 2 panels show the PD-1 KO ILC2 activation trajectory. The lines correspond to the principal graph learned by Monocle 3