Histological Assessment of Cardiac Injury

Tissues from rats were fixed using 10% buffered formalin, dehydrated, embedded in paraffin, and sectioned into 5 μm-thick sections. Hematoxylin and eosin (HE) staining was used to assess cardiac injury, whereas Masson’s trichrome staining was used to detect collagen fibers, and the slides were observed under an optical microscope. For immunohistochemistry, sections were stained with a primary antibody against SGLT1 (1:200, Abcam) and then stained with a secondary antibody. After washing with PBS, the slides were incubated with 3,3′-diaminobenzidine. The detailed procedure has been described previously (Lin et al., 2019 (link)). Semiquantitative analysis was performed using image analysis software (Image-Pro Plus, Media Cybernetics).