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Sections were washed twice in TBST, followed by detection of secondary antibody using chromogen substrate buffer DAB+ (#K3468, Dako North America, Inc., Carpinteria, CA, USA) and DAB-chromogen (#K3468, Dako North America, Inc., Carpinteria, CA, USA)

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For PAS staining to evaluate glycogen and basement membranes, tissue sections were oxidized with periodic acid (C0142S, Beyotime Biotechnology, China), treated with Schiffs reagent (C0142S, Beyotime Biotechnology, China), and counterstained with Hematoxylin (Bry-0001-01, Runner bio, China)