Phospho-DDIT3/CHOP (Ser30) Antibody - #AF3277

Fig. 7. p53-induced TRIB3 upregulation depends on the transcription factor CHOP. (A) The mRNA level of Trib3 was analyzed by qRT-PCR in H9c2 cells (n = 3). (B) Transcription factors were predicted to bind to Trib3 by FIMO. Predictive intersection of rat and mouse. Screening condition q-value < 0.05. (C) CHOP binding motif provided by the JASPAR database. Cell-based luciferase reporter assays were detected in H9c2 cells co-transfected with luciferase-expressing vectors driven by the Trib3 promoter or the mutant promoters, together with an empty plasmid as the control for 24 h (n = 3). (D, E) ChIP analysis was performed with anti-CHOP antibody or IgG in H9c2 cells. The precipitated chromatin was analyzed by qRT-PCR (n = 3). (F) Proteins interacting between p53 and CHOP were retrieved from the STRING database ( https://string-db.org/ ). (G) Interaction of p53 and CHOP in H9c2 cells determined by co-IP. (H) Co-localization of p53 and CHOP was determined by immunofluorescent staining in H9c2 cells. (I) The expression of p-CHOP and CHOP in cardiac tissues was examined by western blot (n = 6). (J) Interaction of CHOP and AMPKα in H9c2 cells demonstrated by co-IP. (K, L) The p-CHOP and CHOP level was detected by western blot analysis after transfection with NC-shRNA or Ampk-shRNA in H9c2 cells (n = 3). (M) CHOP was immunoprecipitated from H9c2 cells of different groups and measured using western blot analysis with an anti-ubiquitin antibody. (N) The immunoblotting of immunoprecipitated CHOP with antibody-recognized ubiquitin of different groups in H9c2 cells. (O) ChIP analysis was performed with anti-CHOP antibody or IgG in H9c2 cells. The precipitated chromatin was detected by qRT-PCR (n = 3). (P) Diagram showing the experimental design for the mice study. (Q) Representative M-mode echocardiographic images. (R) Western blot analysis of p-AKT, p-GSK-3β, and p-GS in cardiac tissues (n = 6). Data shown in the graphs represents the means ± SD. *P < 0.05; N.S., not significant.