Transcriptome-wide RNA-seq, m⁶A-seq, and RIP-seq assays in murine NK cells. (A) Volcano plots of differentially expressed genes from RNA-seq. (B) GSEA showing enrichment of E2F targets, G2/M checkpoint, and mitotic spindle hallmark gene sets in Ythdf2^ΔNK NK cells compared with Ythdf2^WT NK cells. (C) The proportion of m⁶A peak distribution in NK cells from Ythdf2^WT and Ythdf2^ΔNK mice. (D) GO analysis of transcripts with m⁶A peaks. (E) Density distribution of the YTHDF2-binding sites across mRNA transcriptome from RIP-seq data. (F) The proportion of YTHDF2 binding site distribution from RIP-seq data. (G) Top 10 GO clusters from GO analysis of YTHDF2 target genes from RIP-seq data. (H) Immunoblotting showing the protein levels of MDM2, TDP-43, and YTHDF2 in IL-2–expanded NK cells enriched from the spleen of Ythdf2^WT and Ythdf2^ΔNK mice. (I and J) qPCR and immunoblotting showing the expression of Mdm2 (I) and Tardbp (J) in NK cells transfected with gene-specific or control siRNA. (K and L) IL-2–expanded NK cells were transfected with Mdm2-specific siRNA cells under the stimulation of IL-15. 3 d later, cell proliferation and apoptosis were analyzed by Ki67 staining (K) and annexin V staining (L), respectively (n = 3 per group). Data are shown as mean ± SD and were analyzed by unpaired two-tailed t test (I and J) or one-way ANOVA with Šídák post-test (K and L). Data are representative of at least two independent experiments. ***, P < 0.001. ES, enrichment score; Fc, fold change; FDR, false discovery rate; NES, normalized enrichment score; Rep, repetition.