IR induces YTHDF2 to target Notch signaling in DC. (A) Venn diagram of overlapping genes from mRNA-seq that were upregulated following downregulated following Ythdf2-cKO+IR versus WT+IR and WT+IR versus WT. (B) Heatmap of differentially expressed genes in DCs from the WT+IR and WT group (left), and heatmap of differentially expressed genes in DCs from cKO+IR in comparison with the WT+IR group (right). (C) Schematic illustration of the Notch signaling pathway. Ligands in signal-sending cells can bind to glycosylated NOTCH receptors on signal-receiving cells. ADAMs are recruited for cleavage of the outside domain of Notch. After cleavage, the remaining part of the NOTCH receptor can be further cleaved on the cell membrane by γ-secretase and transported into lysosomes for generation of NICD. NICD can be translocated into the nucleus to crosstalk with other signaling pathways and regulate transcription. (D) Enrichment of Mfng, Aph-1b, and Aph-1c mRNA in the YTHDF2-immunoprecipitated RNA fraction of BMDCs, determined by RIP-qPCR (n = 3). (E–G) BMDCs from WT and Ythdf2-cKO mice were treated with actinomycin D. mRNA was collected at indicated time points after treatment, and mRNA levels of (E) Aph-1b, (F) Aph-1c, and (G) Mfng were measured by RT-qPCR (n = 3). (H) mRNA level of Mfng, Aph-1b, and Aph-1c from tumor-infiltrating DCs of WT, WT+IR and cKO, and cKO+IR mice (n = 3). (I) mRNA expression level of Notch receptors in WT or cKO DCs that were cocultured with irradiated tumor cells (n = 3). (J) mRNA expression level of Notch ligands in tumor cells with or without irradiation (n = 3). Statistical analysis was performed using two-sided unpaired Student’s t test (C–H); **P < 0.01; ***P < 0.001. Data are represented as the mean ± SEM. ADAMs, a disintegrin and metalloproteases.