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. 2021 Mar 19;12(10):3067–3076. doi: 10.7150/jca.54759

Figure 5.

Figure 5

ZEB1 is a direct target of miR-369 in HCC cells. A. A potential target site for miR-369 in the 3'-UTR of human ZEB1 mRNA, as predicted by the program Targetscan. To disrupt the interaction between miR-369 and ZEB1 mRNA, the target site was mutated. B. Luciferase reporter assays performed in HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells transfected with wild-type or mutant ZEB1 3'-UTR constructs. C. The mRNA expression of ZEB1 was checked in HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells by real-time PCR. D. The protein expression of ZEB1 was checked in HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells by western blot. GAPDH was used as an invariant control. E. The protein expression of ZEB1 was checked in HCCLM3 miR-369 sponge or Huh7 miR-369 sponge and their control cells by western blot. GAPDH was used as an invariant control. F. Significant correlation was observed between miR-369 and ZEB1 expression in human HCC tissues (n=20). G. HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells were transfected with ZEB1 siRNA and then checked by western bolt assay. GAPDH was used as an invariant control. H. HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells were transfected ZEB1 siRNA and then subjected to CCK8 assay. I. HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells were transfected ZEB1 siRNA and then subjected to migration assay. J. HCCLM3 miR-369 mimic or Huh7 miR-369 mimic and their control cells were transfected ZEB1 siRNA and then subjected to Invasion assay.