Journal: Arthritis Research & Therapy
Article Title: Increased neutrophil extracellular traps activate NLRP3 and inflammatory macrophages in adult-onset Still’s disease
Figure Lengend Snippet: AOSD NETs are potent triggers of NLRP3 inflammasomes. After stimulation with neutrophil extracellular trap (NET) DNA from adult-onset Still’s disease (AOSD) and healthy controls (HC), mitochondrial DNA (mtDNA), genomic DNA g(DNA), and RNA, the expression ( a ) and secretion ( b ) of interleukin (IL)-1β and IL-18 in THP-1 monocytes were measured using RT-PCR and ELISA, respectively. c Representative immunoblot analysis for NLRP3 inflammasomes in THP-1 monocytes. The images are representative of three independent Western blot experiments. d After stimulation with NET DNA and non-NET source nucleic acids shown above, the expression of IL-1β and IL-18 in PBMC-derived CD14 + monocytes from healthy controls was measured using RT-PCR. e The expression of IL-1β, pro-IL-1β, and NLRP3 in cell lysates (LYS) and IL-1β in supernatants (SN) from THP-1 monocytes after stimulation with NET-DNA in the presence of the NLRP3 inhibitor MCC950 and DNase I was measured by Western blot. The images are representative of three independent Western blot experiments. The histograms show the means ± SD. * P
Article Snippet: Media were subsequently removed and replaced with phenol red-free, serum-free RPMI prior to treatment with 250 ng NET DNA, mtDNA, genomic DNA, RNA, or 5 mM ATP (Sigma, St. Louis, MO) for 2 h. mtDNA, genomic DNA, and RNA were isolated from 293 T cells according to previously published methods [ ].
Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Derivative Assay