Figure 5.

KDM3A Regulates the Expression of YAP1 and TGF-β2 by Altering Enhancer Activity

(A) The expression of YAP1 and TGF-β2 upon sh-KDM3A transfection detected by qRT-PCR. *p < 0.05 versus cells transfected with sh-NC; ^#p < 0.05 versus AA-incubated cells transfected with sh-NC. (B) Enrichment of KDM3A in the promoter region of YAP1 detected by a ChIP assay. (C) Enrichment of KDM3A in the promoter region of TGF-β2 detected by a ChIP assay. In (B) and (C), *p < 0.05 versus the IgG group; ^#p < 0.05 versus the control + sh-NC group; ^&p < 0.05 versus AA-incubated cells transfected with sh-NC. (D) The effect of enzymatic activity of KDM3A on YAP1 and TGF-β2 expression detected by qRT-PCR. *p < 0.05 versus cells transfected with oe-NC; ^#p < 0.05 versus cells transfected with oe-KDM3A; ^&p < 0.05 versus AA-incubated cells transfected with oe-NC; ^@p < 0.05 versus AA-incubated cells transfected with oe-KDM3A. (E) The enrichment of H3K9me2 in the promoter region of YAP1 and TGF-β2 detected by a ChIP assay. *p < 0.05 versus cells transfected with sh-NC; ^#p < 0.05 versus AA-incubated cells transfected with sh-NC. (F) The effect of UNC0631 on YAP1 and TGF-β2 expression detected by qRT-PCR. *p < 0.05 versus cells transfected with sh-NC; ^#p < 0.05 versus cells transfected with sh-KDM3A and veh; ^&p < 0.05 versus AA-incubated cells transfected with sh-NC; ^@p < 0.05 versus AA-incubated cells transfected with sh-KDM3A and veh. (G) Enrichment of H3K27ac and TEAD1 in the promoter region of TGF-β2H detected by a ChIP assay. (H) The enrichment of TEAD1 and H3K27ac in the enhancer of TGF-β2 detected by a ChIP assay. In (G) and (H), *p < 0.05 versus cells transfected with sh-NC; ^#p < 0.05 versus AA-incubated cells transfected with sh-NC. The above data were all measurement data and are expressed as mean ± standard deviation. Data among multiple groups were analyzed by one-way ANOVA, followed by a Tukey’s multiple comparisons post-test. The cell experiment was conducted in triplicate.