YTHDF2 expression in murine NK cells in response to IL-15 stimulation, MCMV infection, and tumor progression. (A and B) The mRNA expression of m⁶A writers (Mettl3, Mettl14), erasers (Fto, Alkbh5), and readers (Ythdf1, Ythdf2, Ythdf3) in murine NK cells (A) and the mRNA levels of Ythdf2 among immune cell subsets (B) were analyzed using the BioGPS online tool. (C) RNA-seq analysis of the online database GSE106138 showing the expression of m⁶A enzymes and readers in splenic NK cells cultured either in the presence or absence of IL-15 (10 ng/ml) overnight. (D) qPCR showing the expression of m⁶A enzymes and readers in splenic NK cells cultured either in the presence or in the absence of IL-15 (10 ng/ml) overnight. 18s rRNA was used as a housekeeping gene for data normalization. Data are shown as mean ± SD. ***, P < 0.001 by unpaired two-tailed t test. Average values from three replicates were calculated for each sample. The experiment was repeated three times independently. (E and F) Representative immunoblot (E) and quantified protein levels of YTHDF2 normalized to actin (F) in splenic NK cells cultured in the presence or absence of IL-2 (100 U/ml), IL-12 (10 ng/ml), or IL-15 (10 ng/ml) overnight. Untreated NK cells were used as negative control (NC group). Data are shown as mean ± SD and were analyzed by one-way ANOVA with Šídák post-test (*, P < 0.05). (G) RNA-seq analysis of online database GSE113214 showing the expression of m⁶A enzymes and readers in splenic NK cells during the MCMV infection. (H) NK cells were isolated from lung tissues of mice injected with B16-F10 at indicated time points. The expression levels of m⁶A enzymes and readers were examined by qPCR. Heatmap showing the expression m⁶A enzymes and readers in the isolated NK cells after B16F10 injection. 18s rRNA was used as a housekeeping gene for data normalization. The expression of each gene is shown as a fold change from the data collected on day 0. Data are representative of at least two independent experiments.