Fig. 6: m6A modification impairs viral RNA sensing efficacy by RLRs.
a Integrative genomics viewer (IGV) plots the m6A sites and RIG-I and MDA5-binding regions on VSV (+) RNA. RNA-seq data were used as input control. b Venn diagram showing the overlap between high-confidence MDA5 and RIG-I binding clusters. The number of clusters in each category is shown in parenthesis. c Biotin-labeled RNA pull-down and Western blot analysis of Rig-I and Mda5 bindings to RNA oligo with or without single m6A modification (Lower graph). The upper graph indicates predicted structure of VSV (+) RNA: 2912–2974. Three times each experiment was repeated independently with similar results. Histograms show mean relative RNA content pulled down from 3 independent replicates. d Immunofluorescence analysis of HeLa cells increased the co-localization between RIG-I/ MDA5 and dsRNA induced by VSV infection for 8 h. n = 20 cells examined over 2 independent experiments with similar results. e RIP-qPCR analysis of increase of RIG-I and MDA5 binding to VSV (+) RNA (region: 1129–1329 nt, referred to Fig. 5a PAR-CLIP result) after deficient for METTL3 in HeLa cells. Data are representative of 2 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, as determined by two-tailed unpaired Student’s t test (c–e). Error bars represent mean ± SEM.
