Rat anti mouse cd8a

For histology, sections were stained with hematoxylin and eosin (H&E), according to established protocols. For immunofluorescence, the sections were air-dried at room temperature for 15 min and with 4% paraformaldehyde for 10 min. Sections were then rinsed with 1 X PBST (0.25% Triton-X100). Blocking was performed for 30 min at room temperature in 5% normal serum in PBS. Sections were incubated with primary antibodies diluted in PBS for 2 h at room temperature. Sections were washed with 1 X PBS and incubated with secondary antibodies diluted in PBS for 1 h at room temperature. After washing again, sections were mounted with the VECTASHIELD HardSet mounting medium with DAPI (Vector Laboratories, Burlingame, CA) and imaged. Primary antibodies used were: rat anti-mouse Ly6G and rat anti-mouse F4/80 (BioLegend, San Diego, CA), rat anti-mouse CD31, rat anti-mouse CD4, and rat anti-mouse CD8a (BD PharMingen, Mississauga, ON). Alexa Fluor 546, goat anti-rat (Life Technology, Burlington, ON) was used as the secondary antibody. All histology and immunofluorescence images are representative of at least three independent animal experiments.