Research Paper Volume 16, Issue 21 pp 13356—13370

Figure 4. DANCR promoted GSK3β-mediated phosphorylation of RXRA protein in cardiomyocytes. (A) Schematic diagram of the predicted RXRA binding sites in DANCR. (B) RIP assay was used to verify the association of RXRA with DANCR. (C) DANCR overexpression vector (pc-DANCR) and control (vector) were transfected into H9C2 cells. The mRNA expression of RXRA was detected with RT-qPCR. (D) shRNA of DANCR (sh-DANCR) and control (sh-NC) were transfected into H9C2 cells, and the levels of RXRA total protein and p-RXRA protein were measured by Western blotting. (E) The protein expression of p-RXRA in myocardium of mice in the sham and AMI groups was detected with Western blotting. (F) H9C2 cells were transfected with DANCR overexpression vector (pc-DANCR) and control (vector), and then treated with OGD. The protein levels of p-RXRA were detected with Western blotting. (G) Co-IP analysis was used to detect the interaction between RXRA and GSK3β by using an antibody of Anti-RXRA. (H) Co-IP analysis was used to detect the interaction between RXRA and GSK3β by using an antibody of Anti-GSK3β. (I) H9C2 cells were transfected with DANCR overexpression vector (pc-DANCR) and/or GSK3β siRNA (si-GSK3β), and then treated with OGD. The protein level of p-RXRA was detected with Western blotting. ^*P < 0.05, ^**P < 0.01.