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. 2021 Jul 10;23:65–81. doi: 10.1016/j.omto.2021.06.017

Figure 5.

Figure 5

ERα may transcriptionally regulate miR16-5p, and miR16-5p may directly target the 3′ UTR of YAP1-mRNA to suppress its protein expression

(A) Two putative EREs predicted by JASPAR from the miR16-1 promoter. (B) ChIP assay results of ERE1/2 of the miR16-1 promoter in QBC939 cells. (C) Wild-type and mutant pGL3-miR16-1 promoter reporter constructs. (D) Luciferase activity after transfection of wild-type or mutant EREs on miR16-1 promoter reporter construct in HuCCT1 cells (left) transfected with ERα-cDNA (oeERα) or pWPI and QBC939 cells (right) transfected with ERα-shRNA (shERα) or vector pLKO. (E) Sequence alignment of the YAP1 3′ UTR with wild-type versus mutant potential miR16-5p targeting sites. (F) Luciferase reporter activity after transfection of wild-type or mutant YAP1 3′ UTR reporter construct in HuCCT1 cells w/wo oemiR16-5p (left) and QBC939 cells treated w/wo miR16-5p inhibitor (right). For (D) and (F), quantitations are presented as mean ± SD. ∗p < 0.05; NS, not significant.