Fig. 4

circPTN sponges miR-145-5p and miR-330-5p. a. Venn diagram for circInteractome and miRWalk of predicting miRNAs sponged by circPTN. b. Dual-luciferase reporter assay showed that co-transfection of WT and mimic miR-145-5p or mimic miR-330-5p markedly decreased luciferase activity in 293T cells, n = 3, *p < 0.05, t test. c. Results of dual-luciferase reporter assay indicated that luciferase activity did not change in 293T cells when miR-145-5p or miR-330-5p binding sites in circPTN were mutated, n = 3, *p < 0.05, t test. d. Left: Biotin-labeled circPTN or anti-sense RNA in agarose gel; Right: Histogram of qPCR of RNA pull-down assay. Compared with group anti-sense, miR-145-5p and miR-330-5p in group circPTN were significantly abundant, n = 3, *p < 0.05, t test. e. Images of Cy3-labeled probe for circPTN and FAM-labeled probes for miR-145-5p/miR-330-5p indicated that circPTN and miR-145-5p/miR-330-5p co-localized in the cytoplasm; scale bar, 20 μm. f. U251 cells stably transfected EV or circPTN were treated with 2 μg/mL actinomycin D or DMSO at indicated time point; the results suggested that compared with group EV, miR-145-5p and miR-330-5p were significantly less than the level observed in circPTN group, n = 3, *p < 0.05, t test. g. circPTN could rescue the downregulation of SOX9 and ITGA5 by miR-145-5p and miR-330-5p, n = 3, *p < 0.05, t test