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  antibodies (Cell Signaling, MA, USA). Double-fluorescent immunohistochemical willpower of cardiomyocyte apoptosis was carried out with terminal dUTP nick
  end labelling (TUNEL) assay on deparafinized 4 μm thick
  sections utilizing an In Situ Cell Dying Detection Kit, fluorescein as per the package protocol (Roche Diagnostics, Mannheim,
  Germany). The reaction product (violet) was visualized with VIP substrate utilizing the Vector ABC Vectastain Elite Equipment
  (Vectorlabs). Reverse transcription proceeded with 1 μg of complete RNA utilizing iScript cDNA Synthesis Equipment (Bio-Rad Laboratories, Hercules, CA,
  USA). Regional myocardial blood stream, 14 days after ischaemia, was measured with stable gold-labelled microspheres (BioPAL, Worcester, MA, USA) as previously described.
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