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. 2020 Jun 25;11(6):486. doi: 10.1038/s41419-020-2689-4

Fig. 6. lncRNA ENST00000563492 functions as a ceRNA for miR-205-5p to regulate the expression of CDH11 and VEGF.

Fig. 6

a MS2bs-based RIP assay in BMSCs 48 h after transfection with MS2bp-YFP plasmid along with MS2bs-ENST00000563492 or MS2bs-Rluc (control vectors). b Luciferase activity of psiCHECK2-ENST00000563492-wt and psiCHECK2-ENST00000563492-mut upon transfection of NC miRNA mimics or miR-205-5p mimics into HEK293T cells. c RIP assay of the enrichment of Ago2 on lncRNA ENST00000563492, CDH11, and VEGF transcripts relative to IgG in BMSCs transfected with Lenti-Vector or Lenti- ENST00000563492. d RIP assay of the enrichment of Ago2 on lncRNA ENST00000563492, CDH11, and VEGF transcripts relative to IgG in BMSCs transfected with pLKO.1-Vector or pLKO.1-ENST00000563492. e miRNA-RIP assay of the enrichment of lncRNA ENST0000056349, CDH11, and VEGF transcript on miR-205-5p relative to NC miRNA mimics in BMSCs transfected with biotin-miR-con or biotin-miR-205-5p. f Western blot analysis of CDH11 and VEGF expression in BMSCs transfected with miR-205-5p mimics, miR-205-5p inhibitors or NC miRNA mimcs and miRNA inhibitors. g Alizarin Red S staining analysis of BMSCs transfected with miR-205-5p mimics, miR-205-5p inhibitors or NC miRNA mimics and miRNA inhibitors. h Western blot analysis of CDH11 and VEGF expression in BMSCs transfected with Lenti-Vector or Lenti-ENST00000563492 along with Dicer ctrl siRNA or Dicer siRNA. i Western blot analysis of CDH11 and VEGF expression in BMSCs transfected with pLKO.1-Vector or pLKO.1-ENST00000563492 along with NC miRNA inhibitors or miR-205-5p inhibitors. j Alizarin Red S staining analysis of BMSCs transfected with Lenti-Vector or Lenti-ENST00000563492 along with Dicer ctrl siRNA or Dicer siRNA. k Alizarin Red S staining analysis of BMSCs transfected with pLKO.1-Vector or pLKO.1-ENST00000563492 along with NC miRNA inhibitors or miR-205-5p inhibitors. BMSCs isolated from patient no. 1, 2, 5, 12, 10, 14, 19, 20, 21, 22, 23, and 24. All experiments were performed at least three times. The results are presented as the mean ± SD. *p < 0.05, **p < 0.01.