Fitc anti mouse human cd11b

The cell suspension was prepared from mice periodontal tissue, including gingiva, periodontal ligament, and a part of alveolar bone as described previously (Wang L. et al., 2020 (link)). The cell suspension was used for flow cytometry analysis (Mizraji et al., 2013 ). Prior to the test, cells were counted and cell viability was evaluated by the Zombie NIR^TM Fixable Viability Kit (423105,Biolegend, Beijing, China). Cells were stained with 1 μg of PE anti-mouse F4/80 (123110, BioLegend, San Diego, CA, United States) solution, FITC anti-mouse/human CD11b (101206, BioLegend, San Diego, CA, United States), PerCP/Cyanine5.5 anti-mouse CD16/32 (101324, BioLegend, San Diego, CA, United States), APC anti-mouse CD206 (141707, BioLegend, San Diego, CA, United States) or anti-rabbit IgG per 1 × 10⁶ cells for 30 min on ice in the dark. After two times washing with PBS, the cells were resuspended in 300 μl PBS and transferred to flow tubes. Flow cytometry analysis was performed using the FACSAria III Cell Sorter (BD Biosciences, San Jose, CA, United States). The mononuclear cells were sorted using anti-CD11b. From mononuclear cells, macrophages were sorted using anti-F4/80. And from macrophages, M1 and M2 macrophages were sorted using CD16/32 and CD 206, respectively.

Anti-β-actin (1:10,000, BH10D10) was bought from Cell Signaling Technology (Danvers, MA, United States); Anti-NLRP3 (1:1,000. Cryo-2) and Anti-ASC (1:1,000, AL177) were purchased from Adipogen (San Diego, CA, United States); Anti-Caspase-1 (1:1,000, ab179515) and Anti-NEK7 (1:10,000 ab133514) were bought from Abcam (Cambridge, CB2 0AX, United Kingdom); Anti-IL-1β (1:000 AF-401-NA; RRID: AB_416684) was obtained from RD systems (Tustin, CA, United States); the DyLight 488-labeled secondary antibody (1:50, A120-100D2) was purchased from InvivoGen (San Diego, CA, United States); and FITC anti-mouse/human CD11b (101216, 1:500 for flow cytometry) and APC anti-mouse Ly-6G (127614, 1:500 for flow cytometry) were from BioLegend.