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. 2020 Jul 22;6(30):eaba3688. doi: 10.1126/sciadv.aba3688

Fig. 5. Induction of methylation-sensitive ERV triggers cytosolic dsRNA signaling and IFNβ production.

Fig. 5

(A) IFN-β mRNA levels, measured by qRT-PCR, in HUVEC transfected with siRNAs targeting ADA2 and innate sensing molecules (n = 3 technical replicates). (B) Differential expression of LTR-containing ERV genes between siControl and siADA2-transfected HUVEC, measured by polydenylated poly(A+)–enriched RNA-seq (n = 3 biological replicates). Red and blue dots denote ERV significantly up- or down-regulated ≥1.5-fold. (C) mRNA levels of ERVFRD1 and ERVK28, measured by qRT-PCR, in HUVEC supplemented for 24 hours with dIno (500 μM) or pretreated for 30 min with the PNP inhibitor 9-deazaguanine (100 μM) before dIno supplementation for 24 hours (n = 3 technical replicates). (D) mRNA levels of ERV at 24 to 120 hours, measured by qRT-PCR, in siADA2- or siDNMT1-transfected HUVEC (n = 3 technical replicates). (E) mRNA levels of ISG and ERV, measured by qRT-PCR, in siADA2-transfected HUVEC treated with anti–IFN-β–neutralizing antibody (40 U/ml) (n = 3 technical replicates). (F) ERV, IFN-β, CCL5, and CXCL10 mRNA levels, measured by qRT-PCR, in HUVEC transfected with control plasmid or plasmids encoding ERVK28 transcript variants. Values are presented as means ± SD. *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001.