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. 2020 Mar 15;10(10):4395–4409. doi: 10.7150/thno.43239

Figure 5.

Figure 5

circPRRC2A function as a sponge for miR-514a-5p and miR-6776-5p in RCC cells. RNA RIP experiments were performed using an antibody against Ago2 onextracts from ACHN A and Caki-1 B cells. C RNA RIP experiments were performed in ACHN cells using the circPRRC2A probe or a control probe. The enrichment of circPRRC2A and potential target microRNAs were detected by qRT-PCR and normalized relative to the control (one-way analysis of variance, Dunnett's test). D Schematic drawing showing the potential miRNAs that might bind circPRRC2A. E-F Schematic of the predicted miR-514a-5p (L) and miR-6776-5p (R) sites in the circPRRC2A. G Luciferase reporter activity of Luc-circPRRC2A in 293t cells after transfection with miR-514a-5p and miR-6776-5p. H Luciferase reporter activity of Luc-circPRRC2A-WT (wildtype) or Luc-circPRRC2A-Mut (mutant) in ACHN cells after transfection with miR-514a-5p and miR-6776-5p. I Silencing of circPRRC2A did not affect the expression level of miR-514a-5p or miR-6776-5p. J Co-localization between miR-514a-5p, miR-6776-5p and circPRRC2A was detected by FISH assay in ACHN and Caki-1 cells. Nuclei were stained with DAPI. K-L miR-514a-5p and miR-6776-5p did not affect the expression level of circPRRC2A, Student t test. Data indicate mean ± SD of three experiments. *P < 0.05, **P < 0.01.