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. 2021 May 3;9:653207. doi: 10.3389/fcell.2021.653207

FIGURE 3.

FIGURE 3

Circ_0021205 serves as a miR-204-5p sponge. (A) RIP experiments with AGO2 antibody were conducted, the expression of circ_0021205 was detected using qRT-PCR. (B,C) The expression of miRNAs bind to circ_0021205 probes in HuCCT1 and KMBC cells was detected by qRT-PCR. (D) Schematic illustration of circ_0021205-WT and circ_0021205-MuT luciferase reporter vectors and the binding sites between circ_0021205 and miR-204-5p. (E) The relative luciferase activities were detected in HuCCT1 and KMBC cells after co-transfection with circ_0021205-WT/MuT and miR-204-5p mimics/NC-mimics, respectively. (F) The expression of miR-204-5p was detected in HuCCT1 and KMBC cells transfected with si-circ_0021205 and OE-circ_0021205 plasmids, respectively. (G) Relative expression of miR-204-5p was detected in CCA and normal tissues. (H) The levels of proliferation makers, Ki-67 and PCNA, were investigated using western blotting. (I) The proliferation of transfected CCA cells was determined by CCK-8 assay. All experiments were repeated at least three time. **P < 0.01, ***P < 0.001.