For histopathological examination, the tissue were fixed with 4% paraformaldehyde (PFA) and embedded in paraffin (JUNSEI, Cat. #58290-1501) using standard procedures. Sections (3 μm) were stained with Hematoxylin & Eosin.
For ANXA10 immunostaining, cattle abomasum samples were collected into cryotube and snap frozen with liquid nitrogen. Tissues were trimmed using cryostat blade (Leica, Cat. #14035838926) and then embedded in Tissue-Tek O.C.T. compound (Sakura, Cat. #4583) and frozen. Section (15 μm) was fixed in 4% PFA at 4 °C for 15 min. Mice stomach tissue were fixed with 4% PFA, and the tissues were rinsed with PBS, 30% sucrose/PBS and embedded in O.C.T compound. Sections (20 μm) were washed with 0.05% Triton x100 (Wako, Cat. #168-11805) in PBS. For ANXA10 immunostaining, the sections were immunostained with anti-Annxin A10 antibody (R&D, Cat. #AF3544, 5 μg/ml) in Canget signal immunostain solution A (TOYOBO, Cat. #NKB-401) for 1 h. For platelet thrombi, the sections were immunostained with anti-integrin beta 3 [EPR2417Y] (abcam, Cat. #ab75872, 1:250) [40 (link)].
For ANXA10 immunostaining, cattle abomasum samples were collected into cryotube and snap frozen with liquid nitrogen. Tissues were trimmed using cryostat blade (Leica, Cat. #14035838926) and then embedded in Tissue-Tek O.C.T. compound (Sakura, Cat. #4583) and frozen. Section (15 μm) was fixed in 4% PFA at 4 °C for 15 min. Mice stomach tissue were fixed with 4% PFA, and the tissues were rinsed with PBS, 30% sucrose/PBS and embedded in O.C.T compound. Sections (20 μm) were washed with 0.05% Triton x100 (Wako, Cat. #168-11805) in PBS. For ANXA10 immunostaining, the sections were immunostained with anti-Annxin A10 antibody (R&D, Cat. #AF3544, 5 μg/ml) in Canget signal immunostain solution A (TOYOBO, Cat. #NKB-401) for 1 h. For platelet thrombi, the sections were immunostained with anti-integrin beta 3 [EPR2417Y] (abcam, Cat. #ab75872, 1:250) [40 (link)].
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