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. 2020 May 30;20(2):1423–1431. doi: 10.3892/ol.2020.11693

Figure 3.

Figure 3.

IGF2BP3 decreases ZO-1 expression by synergizing with miR191-5p via the RISC. (A) ZO-1 mRNA expression was detected in the argonaute 2 complex using an RIP assay in HA22T cells transfected with pLKO and shIGF2BP3. (B) Reverse transcription-quantitative PCR was used to screen 6 potential miRNAs which may be able to regulate ZO-1 from the argonaute 2 complex in Huh-7 cells transfected with pWPI and IGF2BP3-cDNA. (C) Transwell invasion assays were performed using Huh-7 cells transfected with pWPI+pWPI, pWPI+oeIGF2BP3, oemiR-191-5p+pWPI or oemiR-191-5p+oeIGF2BP3. (D) Western blotting was performed to detect ZO-1 expression in Huh-7 cells transfected with pWPI+pWPI, pWPI+oeIGF2BP3, oemiR-191-5p+ pWPI or oemiR-191-5p+oeIGF2BP3. For C and D, the quantifications are shown on the right, and are presented as the mean ± SD. *P≤0.05 and **P≤0.01. ns, not significant; IGF2BP3, insulin-like growth factor II mRNA-binding protein 3; ZO-1, zonula occludens-1; miRNA/miR, microRNA; RISC, RNA-induced silencing complex; RIP, RNA interaction-precipitation; sh, short hairpin (RNA); oe, overexpression; NC, negative control.