Rat anti f4 80

Paraffin-embedded tissue sections (4 μm) underwent appropriate heat-induced antigen retrieval according to standard protocols. Samples were then treated with 3% hydrogen peroxide for 10 min, blocked and incubated overnight at 4°C with murine anti-F. nucleatum (1:10, prepared by our laboratory bacterial immunized mice), rabbit anti-Arg1 (Proteintech, Wuhan, China), rabbit anti-iNOS (Proteintech, Wuhan, China), rat anti-F4/80 (Proteintech, Wuhan, China), rabbit anti-PI3K (Abcam, Cambridge, MA, USA) and mouse anti-AKT2 (Abcam, Cambridge, MA, USA). At the same time, we set up a control by selecting serum from the same animal of the first antibody instead of the first antibody in each group of experiments. The next day samples were washed prior to incubation with secondary antibodies against mouse or rabbit IgG conjugated to HRP (Zhongshang Goldenbridge), and a 5–10 min incubation with 3,3′-diaminobenzidine tetrachloride was used to visualize staining via light microscopy (Olympus, Japan).