Figure 7: circHIPK3 sponges with miR-124 and inhibits its activity.

(a) Proliferation assessed using a CCK-8 kit in HEK-293 T cells transfected with nine miRNA mimics or control RNA (20 nM). (b) qRT–PCR analysis of circHIPK3 level in the streptavidin captured fractions from the HEK-293 T cell lysates after transfection with 3′-end biotinylated miR-124 or control RNA (NC). (c) Co-localization between miR-124 and circHIPK3 was observed (arrowheads) by RNA in situ hybridization in HeLa cells after co-transfection with circHIPK3 and miR-124 expressing vectors. Nuclei were stained with DAPI. Scale bar, 5μm. (d) qRT–PCR analysis of IL6R and DLX2 expression in HEK-293 T cells after transfected with si-cHIPK3, miR-124 mimics or miR-124 with circHIPK3 expressing vector (p-cHIPK3). (e) Proliferation assessed using a CCK-8 kit in cells transfected with circHIPK3 or miR-124 (10 nM) as indicated. Data in a,b,d are the means±s.e.m. of three experiments. (f) qRT–PCR for the abundance of circHIPK3 relative to ACTB and miR-124 relative RNU6B in six human normal tissues. The correlation between circHIPK3 and miR-124 is also shown. *P<0.05, **P<0.01 (Student’s t-test).