Extended Data Fig. 3: The cooperative effects between IFNα and SLE IgG on cell death.
From: Glutathione peroxidase 4–regulated neutrophil ferroptosis induces systemic autoimmunity
a-c. HC neutrophils were cultured in the presence of HC or SLE serum with or without the addition of Cl-amidine (Cl) (peptidyl arginine deiminase 4 (PAD4) inhibitor, 100 μM), or LPX-1 (1 μM) for 4 or 16 hours and NETs were assessed in SYTOX Green+ cells based on morphology (n = 6). Neutrophils with DNA area greater than 400um2 were considered as NETs. Dot plots show the percentage of cells forming NETs in all dead neutrophils from the indicated group. d-e. Representative fluorescent images and related quantification of NETosis. HC neutrophils (n = 6) were stimulated by PMA (50 nM) with or without LPX-1(1 μM) for 4 hours. f-i. HC neutrophils were cultured with SLE IgG (3.6 g L-1) and/or IFN-α (10^5 U ml-1) for 4 or 16 hours and cells were stained with SYTOX Green for the detection of NETs. Dot plots show the immunofluorescence microscope quantification of NETosis in total dead neutrophils from the indicated group (4 h: n = 6; 16 h: n = 3). The scale bar represents 50 μm. Data are shown as mean ± SD. ns p > 0.05. Two-tailed paired Student’s t-test was applied.
