Fig. 3. RNA-LL37 trigger the release of NETs containing further RNA, DNA, and LL37.

a EM pictures from PMNs stimulated with RNA-LL37 (n = 2, scale bar = 2 µm). b Neutrophil elastase (NE) release from PMNs stimulated for 3 h (n = 8, each dot represents one donor). Fluorescence microscopy of fixed and Hoechst- anti-LL37- SYTO RNAselect- (c) and/or anti-ΨU-stained (d, with or without RNase A treatment) or live (e) PMNs stimulated as indicated (n = 6, scale bar = 10 µm or n = 4, scale bar = 20 µm). f Quantification of live-cell microscopy shown in Supplementary Movies 1–3. g, h Skin sections from healthy (g) or psoriasis- (g, h) affected skin (n = 12 patients and three healthy controls, scale bar = 20 µm). i, j RNA-LL37-induced PMN influx (i, n = 3 each, measured by GFP in vivo fluorescence) and ear swelling (j, n = 6 each, mm × 0.01) in LysM^EGFP/+ mice. k Ear sections from RNA-LL37 and PBS injected mice stained for ΨU, citH3 and MPO (n = 6, scale bar = 20 µm). l PMNs stimulated with transferred NETs (n = 3). m Quantification of (l). AF = autofluorescence. (b), (f), (i), (j), and (m) represent combined data (mean + SD) from ‘n’ biological replicates. In (a), (c–h) and (k, l) representative samples of ‘n’ replicates or donors are shown. Arrowheads indicate released RNA/NETs (c–e) or RNA-LL37 co-localization (h). *p < 0.05 according to one-way (b) or two-way ANOVA (f, i, j) with Dunnett’s correction or Kruskal–Wallis test with Dunn’s correction (m). Source data is provided as a Source data file.