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. 2021 Mar 19;12(4):297. doi: 10.1038/s41419-021-03566-2

Fig. 5. CircSPI1 interacts with microRNAs to regulate apoptosis functional signaling pathways.

Fig. 5

A The subcellular localization of circSPI1 was analyzed by RT-qPCR. B Potential targeted miRNAs of circSPI1 were predicted by the circBank and Interactome databases. C Luciferase activity assay detected the miRNAs bound by circSPI1. HEK-293T cells were transfected with the circSPI1 luciferase reporter plasmid and predicted miRNA mimics. The luciferase activity was detected 24 h after transfection. D Luciferase activity of mutant circSPI1 was analyzed in HEK-293T cells transfected with the three miRNA mimics. The schematic diagram delineated putative binding sites of miR-1307-3p, miR-382-5p, and miR-767-5p on circSPI1 (left two panels). The luciferase activity was detected 24 h after transfection (right panel). E The expression levels of the three miRNAs were detected with or without circSPI1 knockdown by RT-qPCR. F Enriched KEGG terms for the three miRNA-regulated genes. G Global view of the circSPI1 and three miRNAs-associated ceRNA network. H The protein levels of apoptosis-related factors were detected with or without circSPI1 knockdown by western blotting. The original gels and three replicates are presented in Supplementary Fig. 4. All bar graphs represent the average of three independent replicates and error bars are SD, *P < 0.05, **P < 0.01, ***P < 0.001, ns no significance.