Cd20 antibody

IHC and IF were performed following standard procedures. Five-micron thick formalin-fixed paraffin embedded (FFPE) human tissue sections were used for the experiments. Stained slides were digitized using the Pannoramic DESK (3D HISTECH) with a 40× objective lens.
For IF, sections were stained with the Djc9 antibody (#NBP1–87903, Novusbio, 1:50) and CD20 antibody (#GB14030, Servicebio, 1:200) following the manufacturer’s instructions. Mean fluorescent intensity (MFI) was quantified using ImageJ (https://imagej.nih.gov/ij/). Colocalization analysis was performed using ImageJ plugin JACoP.
For IHC, sections were stained with the Djc9 antibody (#NBP1–87903, Novusbio, 1:500). Staining intensity score, nuclear staining intensity score, and staining percentage were quantified using ImageJ IHC Profiler plugin. Staining index was calculated as follows: staining intensity score (0–3) × staining percentage (0–100%). Staining intensity score was graded as follows: score 0: negative staining, score 1: weak staining, score 2: moderate staining, score 3: strong staining.

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.