Extended Data Fig. 7: TLR3 and TLR7 sense de-N-glycosylated glycoRNAs.
From: RNA N-glycosylation enables immune evasion and homeostatic efferocytosis
(A) IFN-β ELISA of supernatants from WT, TLR3 KO, and TLR7 KO peritoneal macrophages stimulated for 24 hours with small RNAs from HeLa cells that were treated with or without PNGase F or in combination with RNase and then column purified to remove the associated N-glycans and enzymes. Poly(I:C), CL097, R837 and LPS were used as controls. (B) TNF and IL-6 ELISA of supernatants from THP1 macrophages stimulated with small RNAs treated with or without PNGase F, or in combination with RNase, unmodified or acp3U-containing synthetic RNAs, as well as poly(I:C), CL097, ssRNA40, 2’,3’-cGMP, R837, and TL8. (C) IFN-β ELISA of supernatants of RAW macrophages 24 hours post-electroporation with free uridine, acp3U, uridine+acp3U in combination or the synthetic 18-nucleotide unmodified or acp3U containing RNAs and poly(dA:dT). Pooled data from 3 independent experiments (A–C) are shown. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, not significant. Two-way ANOVA with Tukey’s test (A) or One-way ANOVA with Šídák’s test (B–C).
