Identification of AGO2 mRNA targets in nuclei of male germ cells by eCLIP. (A) Immunoblot analysis of AGO2 immunoprecipitation. (B) Size selection of cDNA (orange and purple bars) following AGO2 IP, linker (147 nt) ligation, and PCR amplification. (C) Overlap of AGO2 targets from two eCLIP replicates from meiotic (left) and postmeiotic (center) cells, and transcripts identified in both replicates of both cell types (right). (D) GO analysis of meiotic consensus targets. (E) Genomic distribution of all AGO2 eCLIP binding regions. (F) Distribution of categories of bound transcript in both cell types (common), meiotic cells only, or postmeiotic cells only. Shuffled control distributions were generated by randomly shuffling a representative CLIP peak set across all annotated transcripts. (TEC) To be experimentally confirmed. (G) Overlap between cytoplasmic and nuclear AGO2 eCLIP targets.
