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. 2021 Mar 18;17(3):e1009438. doi: 10.1371/journal.ppat.1009438

Fig 4. circDtx1 functions as a miRNA sponge of miR-15a-5p.

Fig 4

(A) Schematic illustration of circDtx1-wt and circDtx1-mut luciferase reporter vectors. (B) The relative luciferase activities were detected in EPC cells after cotransfection with circDtx1-wt or circDtx1-mut and mimics or NC. (C and D) The concentration gradient (C) and time gradient (D) experiment of miR-15a-5p mimics was conducted. (E and F) circDtx1 downregulated GFP expression. EPC cells were cotransfected with circDtx1-wt or circDtx1-mut and mimics or NC. The fluorescence intensity and the GFP expression were evaluated by enzyme-labeled instrument and western blotting, respectively. (G) The Ago2-RIP assay was executed in MIC cells after transfection with miR-15a-5p, miR-15a-5p-mut, and NC, followed by qPCR to detect cirDtx1 expression levels. (H and I) RNA pull-down assay was executed in MIC cells, followed by qPCR to detect the enrichment of circDtx1 and miR-15a-5p. (J) The MS2-RIP assay was executed in MIC cells after transfection with pLC5-circ-MS2, pLC5-MS2-circDtx1, pLC5-MS2-circDtx1-mut, followed by qPCR to detect the enrichment of miR-15a-5p. All data represented the mean ± SD from three independent triplicated experiments. *, p < 0.05; **, p < 0.01.