Journal: Ecotoxicology and environmental safety

Article Title: Mechanistic insights into severe pulmonary inflammation caused by silica stimulation: The role of macrophage pyroptosis.

doi: 10.1016/j.ecoenv.2023.114975

Figure Lengend Snippet: Fig. 2. Silica exposure induces mitochondrial depolarization and disrupts energy homeostasis in RAW-ASC cells. RAW-ASC cells were primed with LPS for 6 h then treated with nano-silica, micro-silica, and ATP for 4 h, respectively. (A) Representative images of JC-1 staining showing the alteration of mitochondrial membrane potential between groups. Red, JC-1 aggregates; Green, JC-1 monomers. Scale bar = 50 µm. (B) Quantitative analysis and comparison of fluorescence intensity in A between groups (n = 6). (C) Measurement and comparison of intracellular ROS content between groups (n = 6). (D) Measurement and comparison of intracellular ATP content between groups (n = 4). ns, not significant, *P < 0.05, **P < 0.01.

Article Snippet: The mouse macrophage cell line RAW264.7 cells stably expressing ASC (RAW-ASC) were obtained from Invivogen (San Diego, CA, USA); Dulbecco’s modified Eagle’s medium of high glucose (DMEM), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody, and goat antirabbit IgG-HRP conjugated secondary antibody were obtained from Servicebio (Wuhan, China); Silica nanoparticles, penicillinstreptomycin, mitochondrial Membrane Potential Assay Kit with JC-1, and adenosine triphosphate (ATP) were purchased from Solarbio (Beijing, China); Brilliant blue G (BBG), lipopolysaccharides (LPS), reactive oxygen assay kit, calcium content chromogenic assay kit, ATP assay kit, and Calcein/PI cell assay kit were purchased from Beyotime (Shanghai, China); KCl was obtained from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China); Fetal bovine serum (FBS) was obtained from BI (Israel); Silica particles of micro-size, and fast green FCF were obtained from Sigma-Aldrich (St. Louis, MO, USA); Primary antibodies of NLRP3, P2X7, Pannexin-1, and IL-1β were purchased from ABclonal (Wuhan, China).

Techniques: Staining, Membrane, Comparison, Fluorescence