Reduced NLRP3 Gene Expression Limits the IL-1β Cleavage via Inflammasome in Monocytes from Severely Injured Trauma Patients

Figure 2

Transfection of isolated CD14⁺ cells with the NLRP3 plasmid increases NLRP3 gene and protein expression. (a) Immunohistological staining of human CD14⁺ monocytes isolated from healthy volunteers (HV). Left panel presents nontransfected cells (ctrl), middle panel displays CD14⁺ cells transfected with plasmid control (PC), while the right panel shows cells transfected with the NLRP3 plasmid. Representative photos are shown. (b) Isolated CD14⁺ monocytes were transfected with NLRP3 plasmid and either untreated or treated with lipopolysaccharide (LPS). Protein extracts were isolated from the cells and analyzed via western blotting as described in Patients and Methods to detect indicated proteins. Representative blots of four independent experiments are shown. (c) Isolated CD14⁺ monocytes from either HV or TP were transfected with NLRP3-encoding plasmid (NLRP3) or plasmid control (PC) or remained untreated (ctrl). Cells were then stimulated with LPS and gene analysis of NLRP3 was represented. Data are shown as mean ± SEM. versus indicated.

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