F4 80 antibody

At the end of the experiment, mice were sacrificed under deep anesthesia followed by the rapid excision of the heart and peripheral blood. The atria were removed from each heart and fixed in 4% paraformaldehyde (PFA) (Beyotime). Paraffin-embedded atrial sections were stained with hematoxylin-eosin (HE) to evaluate the degree of inflammatory infiltration and myocardial damage under intense inflammation. Sections of atria also were stained with Masson's trichrome to evaluate the distribution and localization of collagen.
Immunohistochemistry (IHC) analysis was performed on paraffin-embedded sections of atria tissues. Paraffin-sections were sequentially subjected to dewax, rehydrate, antigen-repaire, block endogenous peroxidase activity, and serum seal. The primary antibodies used were EGFR antibody (Servicebio, 1:1000), CD3 antibody (Servicebio, 1:300), CD20 antibody (Servicebio, 1:300), F4/80 antibody (Servicebio, 1:700) and Tryptase (Abcam, 1:100) which were added and incubated with sections overnight at 4℃. The corresponding species of primary antibody were added and incubated at room temperature for 50 minutes after three time washing with PBS. After DAB chromogenic reaction, nucleus counterstaining and mounting, images were then obtained using an automatic digital slide scanning system (KFBIO, KF-PRO-120) and analyzed using ImageJ.

Mouse liver tissues were embedded in an optimal cutting temperature (OCT) compound (Servicebio) to make frozen sections. Serial sections of the tissues of 8 μm thickness were cut and used for the fluorescence observations. Double immunofluorescent staining was performed to evaluate immune cell infiltration with Ly6G antibody (Servicebio) and F4/80 antibody (Servicebio). The apoptosis of hepatocytes was detected using a terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay from a commercial kit (Yeasen), following the manufacturer’s instructions. The nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI) (Beyotime, Shanghai, China), and the positive cells were visualized using a fluorescence microscope.

Tumor tissues were collected from MB49-bearing mice intravenously injected with bacteria (1x10⁷ CFU per mice) or PBS for hematoxylin and eosin (H&E) staining. Macrophages were labeled with F-4/80 antibody (Servicebio, GB11027). M1 subset macrophages were marked with iNOS and CD68 antibody (Servicebio, GB11119, GB11067). Neutrophils were labeled with Ly-6G antibody (Servicebio, GB11229). The complement activity was recognized by C3 antibody (Abcam, ab200999). T and B cells were labeled respectively with CD3 antibody (Servicebio, GB13014) and CD19 antibody (Servicebio, GB11061). The collagen was stained with Sirius red staining. The mouse primary antibodies was detected using a goat anti-mouse secondary antibody (Servicebio, gb111739).