Figure 6: circHIPK3 serves as a sponge for multiple miRNAs in human cells. | Nature Communications

Figure 6: circHIPK3 serves as a sponge for multiple miRNAs in human cells.

From: Circular RNA profiling reveals an abundant circHIPK3 that regulates cell growth by sponging multiple miRNAs

Figure 6

(a) Schematic illustration showing that the conservation across 100 vertebrate species and AGO2 binding sites in circHIPK3 genomic region. (b) Ago2 RNA immunoprecipitation (RIP) assay for the amount of circHIPK3 in HEK-293 T cells stably expressing Flag-AGO2 or Flag-GFP. Data are the means±s.e.m. of three experiments. (c) The entire circHIPK3 sequence (red) was cloned into the downstream region of the luciferase gene, denoted LUC-cHIPK3. Luciferase reporter assay for the luciferase activity of LUC-cHIPK3 in HEK-293 T cells co-transfected with siRNA against circHIPK3 or circHIPK3 expressing vector. Data are the means±s.e.m. of three experiments. (d) Luciferase reporter assay for the luciferase activity of LUC-cHIPK3 in HEK-293 T cells transfected with a library of 424 miRNA mimics to identify miRNAs that were able to bind to the circHIPK3 sequence. Nine miRNAs that inhibited luciferase activity by 30% are indicated by red dots. (e) A schematic drawing showing the putative binding sites of the miRNAs associated with circHIPK3. (f) Luciferase reporter assay for the luciferase activity of LUC-cHIPK3 or LUC-cHIPK3-mutant in HEK-293 T cells co-transfected with miRNA mimics. Data are the means±s.e.m. of three experiments.

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