Abundant differential cytosolic mRNA levels and differential translation between NK cells activated with IL-15 or IL-2 following cytokine withdrawal. (A) Overview of gene expression studies. Isolation of polysome-associated mRNA (ie, associated with >2 ribosomes) and cytosolic mRNA from 6 donors and 4 conditions followed by generation of smartSeq2 RNAseq libraries. (B) Relative (IL-15 vs IL-2) polysome-associated mRNA levels for IL-2Rα and CD56 after 48 hours of cytokine activation and following withdrawal for an additional 24 hours. (C) Densities of genome-wide FDRs comparing IL-15– to IL-2–activated cells post-cytokine withdrawal using data from cytosolic or polysome-associated mRNA and translational efficiency as identified by anota. (D) Genome-wide log2 fold changes (IL-15 vs IL-2) following cytokine withdrawal using data from cytosolic or polysome-associated mRNA. mRNAs with differential polysome-association (green) or differential translational efficiency (red; by anota analysis) are indicated. (E) Heatmap showing log2 fold changes using data from cytosolic or polysome-associated mRNA for mRNAs showing differential polysome association between NK cells activated with IL-15 vs IL-2 following cytokine withdrawal. (F) Heatmap showing log2 fold changes using data from polysome-associated mRNA for differentially expressed genes belonging to the indicated gene ontology (GO) processes. (E-F) The sidebar indicates differentially translated genes identified by anota analysis (black).