Fig. 3

CircUCK2(2,3) acts as a sponge of miR-149-5p. A AGO2 RIP assay followed by qRT–PCR showed enrichment of circUCK2(2,3) and circSMARCA5 in AGO2 precipitates. B ChIRP assays demonstrated the enrichment of AGO2 with the circUCK2(2,3) precipitates but not with the linear UCK2. C Combined bioinformatic analysis for putative miRNAs binding with circUCK2(2,3). D ChIRP assays followed with Taqman miRNA assays showed the enrichment of miR-149-5p but not miR-580-3p with the circUCK2(2,3) precipitates. E Dual luciferase assays were performed in 293T cells transfected with miR-149-5p mimics together with reporter vectors carrying WT circUCK2(2,3) or circUCK2(2,3) mutant with mutated miR-149-5p binding site. Statistical analyses were performed using paired Student’s t-tests (NS, no statistical significance; *p < 0.05; ***p < 0.001). AGO2 argonaute RISC catalytic component 2, lin probes linear probes, circ probes circRNA probes, WT wild type, MUT149 mutant with mutated miR-149-5p binding site