Tumors exploit FTO-mediated regulation of glycolytic metabolism to evade immune surveillance

1）We co-cultured OT-1 CD8+ T cells with shFto or shNC B16-OVA melanoma cells. After 8 hr of co-culture, we sorted OTI CD8+ T cells via flow cytometry and performed RNA-seq, in order to examine the impact of tumor-intrinsic FTO on T cells；2）To investigate how Fto-Kd dampened glycolytic metabolism in B16-OVA cells, we performed RNA-seq on Fto-Kd and control tumor cells; 3）ATAC-seq was performed on shFto or shNC B16-OVA melanoma cells to characterize the chromatin accessibility and infer the upstream regulatory factors of differentially expressed genes；4）To identify the target mRNA transcripts of FTO, MeRIP-Seq (m6A-seq) was performed on shFto or shNC B16-OVA melanoma cells with or without Dac51 treatment. B16-OVA cells are a cell line cultured in the laboratory, and OT-1 T cells are sorted from the spleens of OTI mice with a C57BL/6 background.