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. 2021 Jan 6;14:83–95. doi: 10.2147/OTT.S277831

Figure 3.

Figure 3

Circ_0006404 directly targets miR-1299 in PCa cells. (A) Through screening with Circular RNA Interactome database, miR-1299 was predicted as a possible miRNA target of circ_0006404, and the binding sites between miR-1299 and circ_0006404 were shown. The site-directed mutation in circ_0006404 was conducted to verify the target relationship between miR-1299 and circ_0006404, and the mutant type sequence in circ_0006404 was also shown. (B) Dual-luciferase reporter assay was performed to confirm the direct combination between miR-1299 and circ_0006404 in PCa cells. (C) RNA-pull down assay was used to verify the target relationship between miR-1299 and circ_0006404 in PCa cells. (D) RIP assay was conducted to explore if there was spatial interaction between miR-1299 and circ_0006404 in RISC in PCa cells. (E) MiR-1299 level in 30 pairs of PCa tumor tissues (n=30) and adjacent normal tissues (n=30) was examined by qRT-PCR. (F) MiR-1299 level in WPMY-1 and two PCa cell lines (LNCaP-AI and DU145) was measured by qRT-PCR. (G) LNCaP-AI and DU145 cells were transfected with inhibitor NC or miR-1299 inhibitor. The abundance of miR-1299 was examined in transfected PCa cells by qRT-PCR. (H) We transfected si-circ_0006404 alone or together with miR-1299 inhibitor into PCa cells. The expression of miR-1299 was measured in PCa cells by qRT-PCR. *P<0.05.