CML-exposed NK cells demonstrate an immature, proinflammatory gene signature. (A) Schematic of scRNAseq experiment. Control and CML-exposed CD45.1⁺ bone marrow (BM) and spleen NK cells were fluorescence-activated cell-sorted for further 10× workflow and Illumina sequencing. (B) Two-dimensional t-distributed stochastic neighbor embedding (t-SNE) plots of recovered single NK cells colored by condition (left plot), K-means cluster assignment (middle plot), or individual gene expression (right plot). (C) Bar plots of frequencies of each cluster in control and CML-exposed NK cells (left); bar plots of frequencies of control and CML-exposed NK cells in each cluster (right). (D) Volcano plot of DEGs between control and CML-exposed K1+K2 NK cells. (E) Pathways significantly enriched in CML-exposed NK cells. (F) Venn diagram of genes significantly upregulated in CML-exposed NK cells and overlapping across tissue-cluster combinations. K1_Spl, cluster 1 NK cells from spleen; K1_BM, cluster 2 NK cells from bone marrow; similar for cluster 2 (K2). (G) RT-PCR analysis validating overexpression of Cish in immature (left) and CD11b⁺ M1+M2 (right) NK cells sorted from control or CML mice. Median with a range, unpaired 2-tailed t test. (H) NK cell active degranulation score in control vs CML-exposed K1+K2 NK cells. Median, unpaired 2-tailed t test; ∗P ≤ .05; ∗∗P ≤ .01; ∗∗∗P ≤ .001; ∗∗∗∗P ≤ .0001.