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

Figure 6.

Figure 6

circPRRC2A serves as a sponge for miR-514a-5p and miR-6776-5p to increase TRPM3 in RCC. A Secondary structure and sequence conservation predictions of miR-514a-5p and miR-6776-5p. B Venn diagram showing the mutual putative target genes of miR-514a-5p and miR-6776-5p. TRPM3 is a mutual putative target gene of miR-514a-5p and miR-6776-5p, Arraystar Human circRNA Array analysis also showing that TRPM3 is differentially expressed in mRCC (middle, heat map). C Biotinylated RNA pulldown assay demonstrates that the miR-514a-5p or miR-6776-5p-captured fractions distinctly enrich TRPM3. D-E miR-514a-5p or miR-6776-5p mimics could significantly suppress the expression level of TRPM3, and this suppression was reversed after over-expression of circPRRC2A. F Schematic of TRPM3 wild-type (WT) and mutant (Mut) luciferase reporter vectors. G-H Relative luciferase activities were analyzed in 293t cells co-transfected with miR-514a-5p mimics, miR-6776-5p mimics, or miR-NC and WT or Mut luciferase reporter vectors. I Western blot analysis showed sh-circPRRC2A or miR-514a-5p/miR-6776-5p mimics could significantly suppresses the expression level of TRPM3, and the suppression was reversed after over-expression of circPRRC2A. J Relative expression levels of TRPM3 in tumor patients derived from TCGA database. K Western blot analysis was used to detect the TRPM3 and VEGFA protein levels in Xenograft tumor tissues transfected with the sh-circPRRC2A compared with the expression in their corresponding controls. Data indicate mean ± SD. *P < 0.05, **P < 0.01.