Heart slices of all animal models were fixed with 10% formalin and then sectioned at the coronal plane. The sections were deparaffinized and refixed in preheated Bouin’s Solution at 56°C for 15 min. After removing the yellow color from sections with running tap water, these sections were stained in BiebrichScarlet-Acid Fucshin for 5 min and in Phosphotungstic/Phosphomolydic Acid Solution for 5 min. Then the slices were stained with Aniline Blue Solution for 5 min to show the nucleus and mounted with xylene. The stained slices were scanned with a digital slide scanner (Pannoramic SCAN, 3DHISTECH and the blue-colored area were quantified with ImageJ software to analyze the area of fibrosis.

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