|
Figure 5
MiR‐29b‐3p regulated NOTCH2 by directly targeting its 3' UTR. A, The mRNA expression of NOTCH2 was decreased in zebrafish embryos injected with miR‐29b‐3p mimic vs NC mimic from 24 to 72 hpf. B, qRT‐PCR results showed that NOTCH2 mRNA expression was decreased in the miR‐29b‐3p mimic group and increased in the inhibitor group. C, WB results showed that NOTCH2 protein expression was lower in cells transfected with miR‐29b‐3p mimic and higher in cells transfected with miR‐29b‐3p inhibitor. D, Relative quantification of NOTCH2 protein. E, HEK‐293 cells was co‐transfected with psiCHECK‐2‐NOTCH2‐3'UTR‐WT‐Human (100 ng) or psiCHECK‐2‐NOTCH2‐3'UTR‐MUT‐Human (100 ng) and miR‐29b‐3p mimic (20 pmol) or NC mimic (20 pmol). F, H9c2 cells were co‐transfected with psiCHECK‐2‐NOTCH2‐3'UTR‐WT‐Rat (100 ng) or psiCHECK‐2‐NOTCH2‐3'UTR‐MUT‐Rat (100 ng) and miR‐29b‐3p mimic (20 pmol) or NC mimic (20 pmol). G, HL1 cells were co‐transfected with psiCHECK‐2‐NOTCH2‐3'UTR‐WT‐Mouse (100 ng) or psiCHECK‐2‐NOTCH2‐3'UTR‐MUT‐Mouse (100 ng) and miR‐29b‐3p mimic (20 pmol) or NC (20 pmol) mimic. MiR‐29b‐3p decreased the relative luciferase activity in the WT group but had no effect in the MUT group. The data are presented as the mean ± SEM. Statistical significance is shown as *