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. 2022 Sep;32(9):1655–1668. doi: 10.1101/gr.276578.122

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© 2022 Griffin et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see https://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 1. — Nucleus-specific protein function for AGO2 in male germ cells. (A) Western blot for AGO2 protein in whole-cell lysate (WCL), cytoplasmic fractions, or nuclear fractions of meiotic (pachytene spermatocyte, SYCP3⁺) and postmeiotic (round spermatid, ACRV1⁺) male germ cells. Tubulin and histone H3 are found in cytoplasmic and nuclear fractions, respectively. Round spermatids have not yet begun histone-to-protamine exchange and therefore have similar levels of histone H3 compared with meiotic cells. (B) Immunoprecipitation of AGO2 for IP-MS in cytoplasmic and nuclear germ cell fractions validated by western blotting with anti-AGO2 antibody. One percent of total lysate was used as input. IP with mouse IgG is a negative control. (C) Common and compartment-specific AGO2 protein interactors in the nucleus and cytoplasm of wild-type postmeiotic cells. Box shows detected miRNA/siRNA pathway components along with the number of unique peptides identified in each compartment. (D) Statistically enriched Gene Ontology (GO) terms for AGO2 protein interactors in the nuclear compartment.