The antibodies used for immunofluorescence double-staining or triple-staining of mice heart tissue sections in each group are as follows: anti-CD86 antibody (ab119857, Abcam, USA), anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-NLRP3 antibody (ab214185, Abcam, USA), anti-ASC-antibody (sc-514414, Santa Cruz, USA), anti-Caspase-1 antibody (ab1872, Abcam, USA), Alexa Fluor 488 (D001-34, Abcam, USA). DAPI (c1002, Beyotime, China) stained cell nucleus. The brief process is as follows: dewaxing, rehydration, repair, blocking, primary antibody incubation at 4°C overnight, PBS washing, secondary antibody incubation in the dark, DAPI staining for cell nuclei, and mounting.
RAW264.7 macrophages were seeded in confocal dishes, after stimulated administration, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% TritonX-100 for 20 min. Then, cells were blocked with 1% BSA in the 37°C incubator for 1 h. Incubation with primary antibody at 4°C overnight, followed by incubation with the secondary antibody for 1 h at room temperature away from light. Then, wheat germ agglutinin (WGA) (AC15L012, Life-iLab, China), labeled with AF488, stained cell membrane for 20 min. Finally, nuclei were stained with DAPI for 5 min. The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).
RAW264.7 macrophages were seeded in confocal dishes, after stimulated administration, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% TritonX-100 for 20 min. Then, cells were blocked with 1% BSA in the 37°C incubator for 1 h. Incubation with primary antibody at 4°C overnight, followed by incubation with the secondary antibody for 1 h at room temperature away from light. Then, wheat germ agglutinin (WGA) (AC15L012, Life-iLab, China), labeled with AF488, stained cell membrane for 20 min. Finally, nuclei were stained with DAPI for 5 min. The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).