Skip to main content
. Author manuscript; available in PMC: 2016 May 7.
Published in final edited form as: Stem Cells. 2016 Feb 1;34(5):1284–1296. doi: 10.1002/stem.2283

Figure 5.

Figure 5

YAP1 is a direct target of miR-194. (A): Putative miR-194 binding sites within the human YAP1-3′UTR are shown in complementary pairing. (B): YAP1 protein expression levels determined by western blot analysis at day 2 in HepaRG-empty and HepaRG-miR-194 cell lines. Expression of α-tubulin was used as a protein loading control. (C): YAP1 protein expression levels determined by western blot analysis at day 5 and day 10 in hESC-empty and hESC-miR-194 cell lines. (D): HepaRG cells were transiently transfected with negative control miRNA (NC; 100 nM) or miR-194 (50 nM or 100 nM) for 48 h. YAP1 mRNA and protein expression levels determined respectively by quantitative PCR and by western blot analysis in HepaRG cells transfected with negative control miRNA or miR-194. (E) Left panel: Cells were transfected with YAP1-3′UTR reporter constructs (200 ng) in the presence or absence of negative control miRNA (100 nM) or miR-194 (50 nM or 100 nM). Effects of miR-194 on the reporter constructs were determined at 48 h after transfection. Right panel: HepaRG-empty and HepaRG-miR-194 cell lines were transfected with YAP1-3′UTR reporter constructs (200 ng). Relative luciferase activities were measured and calculated as the ratio of firefly/renilla activities in the cells, and normalized to those of the negative control miRNA. The results were presented as means ± SEM from three independent experiments (P < 0.05). (F): HepaRG cells were transiently transfected with negative control miRNA (NC; 100 nM) or antisense miR-194 inhibitor (50 nM or 100 nM) for 48 h. MiR-194 levels determined by quantitative PCR in in HepaRG cells transfected with negative control miRNA or antisense miR-194 inhibitor. (G): YAP1 mRNA and protein expression levels determined respectively by quantitative PCR and by western blot analysis in HepaRG cells transfected with negative control miRNA or antisense miR-194 inhibitor. (H): Cells were transfected with YAP1-3′UTR reporter constructs (200 ng) in the presence or absence of negative control miRNA (100 nM) or antisense miR-194 inhibitor (50 nM or 100 nM). Effects of antisense miR-194 inhibitor on the reporter constructs were determined at 48 h after transfection. Relative luciferase activities were measured and calculated as the ratio of firefly/renilla activities in the cells, and normalized to those of the negative control miRNA. The results were presented as means ± SEM from three independent experiments (P < 0.05).