Fig 1: Increased ZFP36L2 expression in CC tissues was negatively correlated with miR-520d-3p. (A) Representative images of different degrees of ZFP36L2 immunohistochemistry staining (-+, weak staining, + moderate staining, ++ strong staining) (left panel) and the percentage of ZFP36L2 high in CC tissues was significantly higher than that in adjacent normal tissues (right panel). Scale bar = 100 μm. (B) The expression of ZFP36L2 mRNA was detected by reverse transcription quantitative PCR in the 57 patients with CC. The values presented as mean ± SD (n = 3). ***p < 0.001, compared with adjacent tissues; (C) The correlation between the expression of ZFP36L2 and miR-520d-3p was analyzed by Pearson's correlation analysis.
Fig 2: ZFP36L2 was a direct target gene of miR-520d-3p. (A) Complementarity of the 3'-UTR of wild-type (WT) or mutant (MUT) human ZFP36L2 mRNA with the miR-520d-3p seed sequence. (B, C) Relative luciferase activity in HeLa and SiHa cell line co-transfected with NC mimics/miR-520d-3p mimics and ZFP36L2-WT/MUT plasmid. (D) Reverse transcription quantitative PCR analyses of ZFP36L2 mRNA expression in HeLa and SiHa cells following transfection of miR-520d-3p mimics or NC mimics. The values presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01, compared with NC mimics. (E) Western blotting results of ZFP36L2 expression following transfection of NC mimics and miR-520d-3p mimics into HeLa and SiHa cells.
Fig 3: Overexpression of ZFP36L2 rescued the effects of miR-520d-3p on CC cell proliferation and migration. HeLa and SiHa cells were transfected with pcDNA-ZFP36L2 and the miR-520d-3p mimics individually or in combination. (A) The overexpression of ZFP36L2 was confirmed by Western blot analysis in HeLa and SiHa cells transfected with pcDNA-ZFP36L2 individually. (B) ZFP36L2 overexpression reversed the suppressive effect of miR-520d-3p mimics on ZFP36L2 expression. (C, D) Cell proliferation was determined by CCK-8 assay in transfected HeLa and SiHa cells. (E, F) A wound healing assay was utilized to evaluate the migration of transfected HeLa and SiHa cells. The values presented as mean ± SD (n = 3). **p < 0.01, ***p < 0.001, compared with NC mimics + pcDNA; ###p < 0.001, compared with miR-520d-3p mimics + pcDNA; (G) The protein levels of E-cadherin, N-cadherin and Vimentin were detected by Western blot analysis in transfected HeLa and SiHa cells.
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