Figure 6.

Progesterone promotes YAP expression by progesterone receptor‐mediated activation of YAP promoter in CM. A, YAP luciferase reporter activity assay. The firefly luciferase/Renilla luciferase ratio in cell lysates was calculated and expressed as relative luciferase activity (P < .0001 vs Control, n = 5). B, Binding of progesterone receptor on YAP promoter was detected by ChIP‐qPCR. DNA/protein complexes of cells treated with or without progesterone were captured by progesterone receptor antibody, and IgG was used as negative control. The immunoprecipitates were further analysed by quantitative PCR (qPCR) using three pairs of primers for YAP gene promoter (n = 4). ^*** P < .001 vs IgG; ^### P < .001 vs Control. C, EMSA assay. Nuclear extracts of NRVMs were incubated with DyLight 800‐labelled DNA oligonucleotide probes containing YAP promoter fragments to show the binding of probe with the corresponding protein in the nuclear extracts; 200‐fold unlabelled probe as competitor was taken as negative control; progesterone receptor antibody was used to show the mobility super‐shift. CM, cardiomyocyte; YAP, yes‐associated protein