Fig. 5

Genomic mapping of in vivo captured MOV10 CLIP targetome. a Schematic of the HITS-CLIP procedure. HITS-CLIP employs the following steps to achieve target specificity: covalent crosslinking via ultraviolet irradiation (254 nm), disruption of protein-protein association by highly stringent wash, gel separation of protein-RNA ribonucleoproteins (RNPs) followed by membrane transfer, RNA retrieval, and expansion for deep sequencing. b Western blot and autoradiography of MOV10-RNA CLIP complexes. Non-crosslinked testes and IgG CLIP served as negative controls. Three independent libraries were prepared from RNA extracted from gel purified RNPs (marked with red line). c Percentage of CLIP reads mapping to genic and intergenic regions. d Percentage of CLIP reads mapping to genomic repeat sequences. The error bars in panel c and d represent variation among three independent CLIP libraries. e, f Genome browser view of CLIP reads over the genomic window for pre-pachytene piRNA cluster 3 (e) and cluster 5 (f). The main peak on the left of the piRNA cluster 5 likely reflects 3′-UTR of upstream mRNAs