Fig. 3

Identification of downstream miRNA molecules by circGLIS3 pull-down experiment. A. Schematic diagram of the circGLIS3 pull-down method using the MS2-tagging system. B. Confirmation of the overexpression of circGLIS3 using qRT‒PCR. C. PCR and Sanger sequencing were used to verify the circular characteristics. D. Western blot analysis of MS2-CP-Flag pulled down by anti-Flag (up). The enrichment of circGLIS3 in the complex with MS2-CP-Flag was measured by qRT‒PCR (bottom). E. Heatmap of the miRNAs pulled down by circGLIS3. F. Venn diagram shows the intersecting miRNAs pulled down by circGLIS3 and predicted by circBank based on the binding sites to circGLIS3. G. CircGLIS3 mutation and wild-type vector construction. H. A dual-luciferase reporter assay to determine the direct binding between circGLIS3 and miR-1343-3p based on their complementary sequences