Extended Data Fig. 9: IFITM1 expression is negatively regulated by YTHDF3 via RNA degradation.
(a) Fold enrichment of IFITM1 was analyzed by RIP-seq in shLacZ or shYTHDF3-HK1 cells. (b) Fold enrichment of IFITM1 was determined by MeRIP-qPCR in shLacZ or shYTHDF3-HEK293 cells and data was presented relative to B2M (2˗ΔCt). (c) KEGG pathway analysis of the 598 proteins from Fig. 6d. Proteins in the DDX family related to RNA degradation were detected. (d) Lentiviruses encoding shYTHDF3 or shDDX5 and their corresponding negative control lentiviruses (shLacZ + shLacZ) were transfected into HK1 cells, and immunoblotting assay were performed with anti-YTHDF3, DDX5, and IFITM1. (e) Cells from Extended Data Fig. 9d were treated with transcriptional inhibitor ActD, followed by IFITM1 detection using RT-qPCR. (f) Cells from Extended Data Fig. 9d were exposed to EBV-GFP for 3 hours and the remaining extracellular viruses were removed by washing with 1×PBS. EBV copy numbers were then measured by TaqMan-qPCR and results from control groups were taken as 100%. Data are presented as mean values ± s.e.m., n = 3 independent experiments, *p < 0.05, **p < 0.01, two-tailed t-test (a, b, e, f).
