Fig. 2.
IL-6 SRE contains an m6A site that is necessary for viral endonuclease protection. (A) PureCLIP scores of the 3′ UTR IL-6 gene in latent and lytic (48 hpr) iSLK.WT cells. Schematic to the right illustrates an IL-6 gene (its 5′ UTR, coding region "CDS" and 3′ UTR): the DRACH motif identified through m6A-eCLIP is in blue and the methylated adenosine in red. (B) Cells were transfected with WTSRE or mutSRE GFP reporter, and total RNA was harvested 24 h later and subjected to meRIP followed by RT-qPCR using GFP primers. Fold enrichment was determined by calculating the fold change of the IP to control Ct values that were normalized through the input. (C) 293T cells transfected with one of three viral endonucleases, as indicated along with the indicated GFP reporters. RNA was collected and quantified using RT-qPCR. ****P < 0.0001; ns, not significant.