Research / 2022 / Article / Fig 5

Research Article

Downregulation of circ-ZNF609 Promotes Heart Repair by Modulating RNA N6-Methyladenosine-Modified Yap Expression

Figure 5

circ-ZNF609 regulates cardiomyocyte survival via modulating the crosstalk between Hippo-YAP and Akt signaling. (a) Western blot analysis demonstrated the Akt and Erk phosphorylation levels in acute I/R heart treated with AAV9-sh-circ-ZNF609 or scrambled control (; ns: nonstatistically significant; mice/group). (b) Representative images of immunofluorescence staining and quantification of the relative TUNEL-positive NRCMs showed that treatment with the Akt inhibitor MK2206 partially blunt the protective effect of circ-ZNF609 knockdown ( wells/group. Scale μm). (c) Western blot analysis of total YAP in cardiomyocytes after transfection with si-circ-ZNF609 or circ-ZNF609 ( wells/group). (d) Representative images of immunofluorescence staining and quantification of the relative TUNEL-positive NRCMs showed that YAP overexpression could block the proapoptosis effect of circ-ZNF609 ( wells/group, scale μm). (e) Representative images of immunofluorescence staining and quantification of the relative Ki67-positive NRCMs showed that YAP overexpression could significantly blunt the decrease of the proportion of cardiomyocytes with Ki67 after circ-ZNF609 overexpression ( wells/group, scale μm). (f) Forced expression of YAP would blunt the inhibition of Akt signaling after circ-ZNF609 overexpression in the OGD/R-induced NRCM apoptosis model ( wells/group). (g, h) Representative images of immunofluorescence staining and quantification of the relative TUNEL-positive (g) and Ki67-positive (h) NRCMs showed that YAP/TEAD inhibitor verteporfin could blunt the prosurvival effects of circ-ZNF609 knockdown in NRCM ( wells/group, scale μm). NC: siRNA negative control; Plo: circRNA overexpression empty vector Plo-ciR without insert circ-ZNF609 sequence; circ-ZNF609: circ-ZNF609 overexpression construct; Fugw: empty vector without YAP overexpression; YAP OE: YAP overexpression construct; OGD/R: oxygen-glucose deprivation/reperfusion. Data are presented as (a, b, d–h) Two-way ANOVA test; (c) independent-sample -test.