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. 2021 Jan 14;10:563486. doi: 10.3389/fonc.2020.563486

Figure 4.

Figure 4

Validation of ST8SIA4 as a direct target of miR-144-5p and miR-451a. (A) The Venn diagram displays the potential targets of miR-144-5p and miR-451a. (B) The sequences of the potential binding sites between 3’ UTR of ST8SIA4 (ST8SIA4-T1-WT, ST8SIA4-T2-WT) and miR-144-5p, miR-451a. ST8SIA4-T1-MUT and ST8SIA4-T2-MUT are the binding sites-mutated segments. (C) Luciferase activities were measured 24 h after the indicated cells transfected with miR-144-5p, miR-451a mimics or inhibitor and reporter plasmid containing wild type 3’ UTR regions of ST8SIA4 or mutated segments. (D) ST8SIA4 mRNA and protein expression levels in the indicated cholangiocarcinoma cells transfected with miR-144-5p, miR-451a mimics or inhibitor. (E, F) ST8SIA4 mRNA (E) and protein (F) expression levels in paired cholangiocarcinoma tissues and adjacent normal bile duct tissues (n=23) examined by qRT-PCR and western blot, respectively. N, normal tissue. T, CCA tissue. (G) Linear regression analysis of the correlation between ST8SIA4 and miR-144-5p or miR-451a in patient samples (n=98). All experiments were repeated 3 times. Bars represent mean ± S.D. *p < 0.05. **p < 0.01. n.s, non-significant.