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. 2019 Dec;7(23):751. doi: 10.21037/atm.2019.11.90

Figure 5.

Figure 5

ZEB1 was the functional target of miR-194. (A) TargetScan showed that the ZEB1 3' UTR among different species was highly conserved and that a conserved region of the ZEB1 3' UTR was complementary to the seed sequence of miR-194. (B) At 48 h after miR-194 transfection, ZEB1 protein levels were decreased compared with the empty vector group (P=0.0231). (C) Luciferase activity was significantly decreased following co-transfection with psiCHECK2-3'UTR-Zeb1 and agomir-194 in HEK293T cells (P=0.038). (D,E) At 48 h after siZEB1 transfection, knockdown of ZEB1 mRNA attenuated the expression of α-SMA at the protein level. (F) At post-transfection 48 h in ARPE-19, overexpression of miR-194 has no significant effect on ZO1 (P=0.1115) and CDH1(P=0.1053) while the mRNA expression of VTN (P<0.0001), CDH2 (P<0.0001), MMP3 (P<0.0001) and MMP9 (P=0.0102) were significantly decreased, respectively. All cell experiments were repeated at least 3 times. The unpaired t-test and the one-way analysis of variance with Tukey’s honestly significant difference post hoc test were used for statistical analysis. ZEB1, zinc finger E-box binding homeobox 1; ZO1, tight junction protein 1; CDH1, E-cadherin; CDH2, N-cadherin; VTN, vitronectin; MMP3, matrix metallopeptidase 3; MMP9, matrix metallopeptidase 9.