Fig 1: Neutrophil phenotype, cytokines and NETs levels in children with COVID-19 according to disease severity. (a–g) Children with COVID-19 (asymptomatic, mild and moderate) and MIS-C were classified as described in Methods. (a–d) Expression of CD11b, CD66b, LAIR-1 and PD-L1 in gated neutrophils was analyzed by flow cytometry in whole blood samples. (a) CD11b (asymptomatic, n = 13; mild, n = 37; moderate, n = 15 and MIS-C, n = 15); (b) CD66b (asymptomatic, n = 10; mild, n = 17; moderate, n = 8 and MIS-C, n = 19); (c) LAIR-1 (asymptomatic, n = 7; mild, n = 9; moderate, n = 5 and MIS-C, n = 16); (d) PD-L1 (asymptomatic, n = 3; mild, n = 14; moderate, n = 4 and MIS-C, n = 8). (e) Spontaneous production of IL-1-β (left) and IL-8 (right) evaluated in supernatants from unstimulated neutrophils (asymptomatic, n = 10; mild, n = 20; moderate, n = 10 and MIS-C, n = 10). (f) Plasma levels of cell-free DNA (left: asymptomatic, n = 13; mild, n = 24; moderate, n = 12 and MIS-C, n = 17) and citrullinated histones H3 (right: asymptomatic, n = 8; mild, n = 7; moderate, n = 4 and MIS-C, n = 8). (g) Levels of cell-free DNA in the supernatant of unstimulated neutrophils from asymptomatic, n = 7; mild, n = 16; moderate, n = 7 and MIS-C, n = 11. Dotted line depicts the median values found in healthy controls. Median and min to max of n donors are shown in a-g. P values were determined by Kruskal-Wallis test with Dunn's posttest and Mann-Whitney U test: * p < 0.05, ** p < 0.01, *** p < 0.001. Asymptomatic (light green circle), mild (dark green circle), moderate (yellow circle), MIS-C (red circle)(For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.) .
Fig 2: Taxol induced a NOXA and GSDME-dependent pro-inflammatory cytokine secretion.A Quantification of IL-1α, IL-1β, IL-6, IL-8, and CXCL1 secretion in 48 h CM from CTCRISPR, NOXACRISPR-/- and GSDMECRISPR−/− MDA-MB-231 cells treated for 24 h or not with Taxol by multiplex cytometry-based technology. B IL-1b secretion measured by ELISA in CTCRISPR, NOXACRISPR−/− and GSDMECRISPR−/− MDA-MB-231 cells after Taxol treatment or not. C Immunoblot analysis of IL-18 secretion in the supernatants of CTCRISPR, NOXACRISPR−/− and GSDMECRISPR−/− MDA-MB-231 cells treated for 24 h or not with Taxol. All experiments were conducted at least three times in independent experiments, and results are presented as mean values ± SEM when relevant (A, B).
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