Anti-miRs Competitively Inhibit microRNAs in Argonaute Complexes
Figure 1
Anti-miRs specifically associate with the Argonaute in vivo and in vitro, only in the context of the cognate target miRNA.
a) Northern analysis of anti-miR-122 in input lysate and in immunopurified Argonaute-containing complexes (IP) from liver tissue of animals dosed subcutaneously, three days prior, with 1, 3 or 10 mg/kg of the compound. Representative experiment shown from 2 independent experiments. b) Western analysis of the levels of Argonaute associated with biotinylated compounds in liver lysates from wild-type and miR-21-deficient animals. 10,3, or 1 pmoles of the 5′-biotinylated compounds were incubated with liver lysates, purified with streptavidin beads and the associated Argonaute proteins were visualized with an anti-Argonaute antibody. Representative experiment shown from three independent experiments. c) Quantification of Argonaute levels associated with the compounds from panel (b). d) Luminescence levels of a miR-21 probe bound by immunopurified miRNA:Argonaute complexes from wild-type or miR-21-lacking liver extracts (miR-21-/-), in the presence of increasing amounts of anti-miR-21. A miR-122 probe in the presence of increasing amounts of anti-miR-122 was used as a positive control for the assay. Representative experiments shown from 3 independent experiments (n = 3).