Arginase-1 Immunofluorescence Study

The tissue sections were heated into citrate buffer 1 mM, pH 6.0, for 15 min at 100°C for antigen retrieval. Then, a working solution of bovine fetal serum 5% and Triton X 0.3% was added and incubated at 37°C in a humidified box for 15 min to block nonspecific antigens. Sections were then incubated overnight at 4°C with primary Arg-1 IgG antibody (1:500, Santa Cruz, SC18351). Afterward, they were rinsed five times in saline 0.9% for 5 min protected from light. Then, they were incubated at 37°C for 60 min with a secondary antibody (1:1,000, Alexa Fluor 488 goat anti-mouse IgG, Thermo Fischer Scientific). Subsequently, sections were rinsed two times in saline 0.9% for 5 min each. Afterward, the sections were incubated with 4,6 diamidinofenilindol (DAPI, Sigma Aldrich), 5 mg/ml solution, for 30 min. Slides were mounted with glycerol and stored protected from light for later observation under a fluorescence microscope. Representative images of five animals per group are shown.