Fig. 7
RIG-I regulated by ALKBH5 affects IFNα secretion through the IKKε/TBK1/IRF3 pathway. a Gene set enrichment analysis (GSEA) showed the signaling pathways enriched after ALKBH5 silencing. b, c The activity of the MAVS/IKKε/TBK1/IRF3 signaling pathway was detected after RIG-I overexpression or silencing in head and neck squamous cell carcinoma (HNSCC) cell lines. d, e The IFNα concentration was measured using an enzyme-linked immunosorbent assay (ELISA) after ALKBH5 overexpression or silencing for 48 h. f Supernatant IFNα was detected using ELISA after si-heterogeneous nuclear ribonucleoprotein protein (HNRNPC) transfection for 48 h. g The RIG-I protein and IFNα concentrations were analyzed after ALKBH5 and/or HNRNPC transfection for 48 h using immunoblotting and ELISA, respectively. h After si-ALKBH5 transfection and TBK1/IKKε-specific inhibitor (10 μM Bay-985) treatment, RIG-I protein and IFNα concentrations were analyzed using immunoblotting and ELISA, respectively. i SCC7-bearing xenografts were established in immunocompetent C3H mice, and murine IFNα (mIFNα) was administered at the indicated times and doses. j The tumor weight was measured after tumor removal in mice. k The serum IFNα concentration was determined using ELISA after mice were sacrificed. l-o Tumor-infiltrating lymphocytes, including NK, T, DC and macrophage cells, were analyzed by flow cytometry after ALKBH5 silencing and mIFNα injection. Data information: Data are presented as the mean ± S.D. Two-tailed unpaired Student’s t test or one-way ANOVA; *P < 0.05, **P < 0.01