CRT silencing improved FZD-induced injury of mitochondria and cardiomyocytes.

<p>A: When cardiomyocytes were transiently transfected for 48 h, the gene silencing effect of CRT was confirmed by RT-PCR (top panel) and western blot (bottom panel). CRT silencing significantly improved the cell viability (B) and MMP (C) of NRCMs stimulated with 50 µM FZD. D: Post-transfection, cardiomyocytes were exposed to 50 µM FZD for an additional 48 h, and annexin V-FITC based apoptosis assay revealed that CRT silencing attenuated cardiomyocyte apoptosis degree. When transfected NRCMs were stimulated with 50 µM FZD for 24 h, real-time PCR revealed that CRT silencing significantly inhibited the decrease of MnSOD mRNA expression (E), and western blot was used to determine the protein levels of CRT, phosphorylated and total STAT3, and MnSOD in the whole cell lysates. β-actin was used as a loading control. A representative blot is shown for each condition (F). Western blot was used to determine the protein expression of phosphorylated and total STAT3 in mitochondria isolated from NRCMs stimulated with 50 µM FZD post-transfection. COXIV was used as a loading control for the mitochondrial protein. A representative blot is shown for each condition (G). All statistical data were obtained from 3–5 independent experiments, and represented as the mean ± SD. *<i>P</i><0.05.