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. 2021 Dec 17;12:730970. doi: 10.3389/fimmu.2021.730970

Figure 3.

Figure 3

Active NK cell cytokine signaling and elevated levels of IL-15 in the bone marrow of leukemic mice. (A–C) CD27+CD11b— NK cells were sorted from leukemic (FLB1 injected) or control (PBS injected) bone marrows (BMs) and RNA sequencing was performed. Data are representative of three independent experiments (five mice/group/experiment). (A) Enrichment pathway analysis showing up-regulated pathways in NK cells from leukemic mice compared to controls. (B) GSEA enrichment plot for the IL-2-STAT5 signaling pathway. (C) Heatmap representing expression of selected genes (IL-2/STAT5 signaling pathway (Reactome), for all genes: p<0.05, Log Expression>4). Color code corresponds to normalized (by row) minimum and maximum gene expression. (D) The levels of IL-15 in the BM supernatants of leukemic or control mice were measured by ELISA (n=9-10 mice/group). (E–G) Freshly isolated BM cells of control and leukemic mice were stained and analyzed by flow cytometry. (E) Zebra plots show the level of phosphorylation of STAT5 and S6 in representative leukemic or control BM-NK cells. The graphs show the MFI of intracellular pSTAT5 and pS6 by total NK cells (n=8 mice/group in two independent experiments). (F) Zebra plots show the expression of CD69 and CD25 in BM-NK cells from leukemic or control mice. The mean percentages of CD69 expressing or CD25 expressing NK Cells in the BM of leukemic and control mice are represented (n=6-7 mice/group in two independent experiments). (G) MFI of the surface expression of IL-15 receptors by total NK cells and by CD27+CD11b— and CD27+CD11b+ NK cell maturation subsets (n=5-7 mice/group in two independent experiments). The values are presented as the mean +/- SEM. *p < 0.05, **p < 0.01, ***p < 0.001, as determined by Mann-Whitney test.