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. 2016 Sep 12;44(22):10603–10618. doi: 10.1093/nar/gkw788

Figure 4.

Figure 4.

miR-24-3p and miR-24-2-5p target Prmt7 3′UTR and impede the stemness of mESCs. (A) Schematic representation of luciferase reporter constructs containing WT Prmt7-3′UTR or its mutants. (B) Relative luciferase activities of reporter constructs containing Prmt7-3′UTR or its mutants in HEK293T cells after transfection of miR-24-3p and miR-24-2-5p mimics. (C and E) Microscopic and AP staining images of V6.5 mESCs after treatment with miR-24-3p (C) or miR-24-2-5p (E) mimic. V6.5 mESCs were treated with miR-24-3p and miR-24-2-5p mimics and incubated for 2d or 4d. (D and F) Quantitative analysis of Oct4, Nanog, Sox2, Klf4, c-Myc and Prmt7 mRNA levels in V6.5 mESCs after treatment with miR-24-3p (D) or miR-24-2-5p (F) mimic. (G) Quantitative RT-PCR analysis of miR-24-3p and miR-24-2-5p levels in WT and differentiated V6.5 mESCs (EBs + RA). (H and I) The effect of LNA-miRNAs (a type of anti-sense microRNAs) against miR-24-3p or miR-24-2-5p on RA-induced mESC differentiation. mESCs were transfected with LNA-miRNAs (H). Oct4, Nanog, Sox2, c-Myc and Klf4 mRNA levels were measured using quantitative RT-PCR (I). In G−I, mESCs were induced to form EBs for 5 days and then treated with RA for another 5 days. Data are presented as the mean ± SD of three independent experiments. P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***) indicate statistically significant changes.