Fig. 5

siRNA off-targets are enriched for seed complementarity in vitro and in vivo. a Volcano plots depicting global gene expression changes in rat hepatocytes at 24 h after transfection with 10 nM of GalNAc-siRNAs of four different sequences. Enrichment of the seed region complementarity in 3′UTRs is tabulated. N = 3 technical replicates. b Volcano plots depicting global gene expression changes in rat liver at 24 h after subcutaneous administration of GalNAc-siRNAs at 50 mg/kg. Two parent GalNAc-siRNAs and their RNAi-inactive versions blocked with inverted abasic (iB) caps are shown. Enrichment of the seed region complementarity in 3′UTRs is tabulated. Blue points, adjusted p ≤ 0.05; red points, adjusted p > 0.05; N = 3 males (6–8 weeks old) per group. The adjusted p-value for fold change was calculate in DESeq2 using the Wald test with multiple test correction. Seed enrichment p-value was calculated using the Fisher’s exact test. The variance was similar between groups that were statistically compared. iB, inverted abasic; NA, not applicable (p-values could not be calculated due to absence of downregulated genes)