Skip to main content
. 2021 May 21;12:648709. doi: 10.3389/fgene.2021.648709

FIGURE 6.

FIGURE 6

DNMT3A was a target gene for circ_0136474/miR-766-3p. (A) Schematic diagram showing the construction of pGL3 luciferase report vectors carrying WT-DNMT3A 3’UTR or MUT-DNMT3A 3’UTR. (B) Dual-luciferase reporter assay measured the relative luciferase activity of WT-DNMT3A 3’UTR and MUT-DNMT3A 3’UTR vectors with co-transfection with miR-766-3p/NC in CHON-001 cells. (C) RIP assay confirmed the relative RNA enrichments of DNMT3A and miR-766-3p in CHON-001 cells. (D) qPCR detected the relative mRNA expression of DNMT3A, and (E–I) Western blotting examined the relative protein expression of DNMT3A in (D,E) OA cartilages (n = 33) and normal cartilages (n = 33), (F) control CHON-001 cells and IL-1β (10 ng/ml)-induced CHON-001 cells, (G–I) IL-1β (10 ng/ml)-induced CHON-001 cells pretransfected with DNMT3A vector or empty pcDNA vector (vector), miR-766-3p/NC or miR-766-3p combined with DNMT3A or vector, si-circ_0136474/NC or si-circ_0136474 combined with anti-miR-677-3p or anti-miR-NC. *P < 0.05 was determined by unpaired t-test, ordinary one-way ANOVA or two-way ANOVA.