Supplementary interactions drive targeting specificity in vitro. (A) Schematic of Ago2 (blue oval) with miRNA and target sequences indicated. Seed (in miRNA) and seed-complementary (in targets) nucleotides are colored yellow. Supplementary (in miRNA) and supplementary-complementarity (in targets) nucleotides are colored red (v1) or blue (v2). (B) Flowchart of target pull-down assays. (C) Results from pull-down of 100 nM target RNAs by 10 nM Ago2–miR122 (each). (D) Results from pull-down of 10 nM target RNAs by 100 nM Ago2–miR122 (each). (E) Results from pull-down of 100 nM target RNAs by 100 nM Ago2–miR122 (each). Enrichment of target-1 (compared to input) for each pull-down is indicated in panels C–E.
