Immunohistochemical Staining of Tissue Samples

Immunohistochemistry (IHC) staining was conducted as previously reported [22 (link)]. In brief, tissue slices were first deparaffinized and gradually rehydrated using a series of graded ethanol solutions. To facilitate antigen retrieval, the deparaffinized tissue slices were subjected to boiling in 10 mM citrate buffer (pH 6.0) for 20 minutes. Afterwards, the tissue slices were incubated with 3% H₂O₂ solution to quench the activity of endogenous peroxidase. The slices were immersed with the specific primary antibody (1:500 dilution, ab109117, Abcam). Visualization of the antibody-antigen interaction was achieved through DAB, followed by counterstaining with hematoxylin.