Anti cd8a clone 4sm15

All FFPE human skin biopsy samples and murine ear skin samples were sectioned into 2 and 4-μm-thick slides, respectively, and subsequently undergone hematoxylin-eosin (HE) staining. Human FFPE skin samples were stained immunohistologically with anti-human CD8 and anti-human CD4 monoclonal antibodies (clone C8/144B and 4B12, respectively, Nichirei Biosciences) using an automatic slide stainer according to the manufacturer’s instructions. The numbers of epidermal-infiltrating cells per sample (magnification, ×400) were counted. Murine ear FFPE samples were stained immunohistochemically with primary anti-CD3 (clone SP7, diluted 1:100, Abcam) and anti-CD8a (clone 4SM15, diluted 1:400, eBioscience) monoclonal antibodies, fluorescent-labeled secondary antibodies (Alexa Fluor^® 488-labeled goat anti-rabbit IgG, and Alexa Fluor^® 555-labeled goat anti-rat IgG, Abcam), and 4′,6-diamidino-2-phenylindole (DAPI) to detect the nucleus, by standard immunohistochemical staining techniques. A fluorescence microscope (BZ-X700, Keyence) was used for observation and to count the number of infiltrating cells per sample (magnification, ×400).