Immunohistochemical Analysis of Tissue Slices

The slices were treated with H₂O₂, boiled twice, and washed according to reported descriptions [28 (link)]. The slices were incubated overnight with rabbit antibodies, α-SMA (1:800, #A17910, ABclonal), TGF-β1 (1:800, #A7040, ABclonal) and LC3B (1:800, (#MA5-37852, 1:800, ThermoFisher Scientific, China) at 4 °C. After washing, the slices were incubated with the secondary antibody (1:1000, AS029, ABclonal) for 120 min at 37 °C, then stained with diaminobenzidine, dehydrated, cleared and sealed. All analysis of immunohistochemical staining sections was performed using ImageJ software.