Research Paper Volume 13, Issue 16 pp 20569—20584

Figure 5. Cry2 interacted with NPAS2 in cardiomyocyte. (A, B) String database (https://string-db.org/) and BioGRID Database (biogrid.org) were used to predict Cry2 and NPAS2 interaction. (C) The protein level of Cry2 (67kDa) in rat myocardial tissue and H9c2 cells was determined using Western Blot. (D) Co-IP assay was performed with anti-NPAS2 or anti-Crry2 antibody was carried out using extracts prepared from rat myocardial tissue and H9c2 cells. The presence of Cry2 or NPAS2 in these IPs was determined using Western Blot. (E, F) Flow cytometry detected the changes of apoptosis in H9c2 cells and quantified. (G) The protein level of Cleaved-Caspase-3 (17kDa) and Caspase-3 (17kDa) in H9c2 cells was determined using Western Blot. (H) The protein level of NPAS2 (90kDa) in H9c2 cells was determined using Western Blot. (I) Representative photomicrographs of NPAS2 (green) and Cry2 (Red) immunofluorescence in H9c2 cells. DAPI was used to counterstain nuclei. Data are expressed as mean ± SEM (n = 3).