Fig. 3.

Fibroblasts function as DAMP sensors via NLRP3 inflammasome signalling. a–e Primary FVB/n NMFs were incubated for 24 h in control medium or in medium containing one of the following DAMPs: ATP 1 mM, H₂O₂ 300 μM, MSU 5 μg/mL, 5% necrotic fluid (extracted from late-stage PyMT tumours). Nlrp3 a and Il1b b expression in fibroblasts were analysed by qRT-PCR. Data are presented as mean ± s.d.; One-way analysis of variance test followed by Dunnett’s multiple comparisons test. Representative of three independent experiments. c Caspase-1 processing was assessed by western blot of cell lysates with anti-Caspase-1. β-actin was utilised as a loading control. The samples shown are derived from the same experiment. Data are representative of three independent experiments. d ELISA quantification of IL-1β secretion. Data are presented as mean ± s.d. of biological replicates (n = 4 per group); Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Representative of three independent experiments. e Primary FVB/n NMFs were incubated for 24 h with 5% necrotic fluid or control medium and expression of selected genes was analysed using qRT-PCR. Data are presented as mean ± s.d. of biological replicates (n = 3 per group); Welch’s t-test. Representative of two independent experiments. f Schematic illustration of cutaneous wound assay. Cutaneous wounds were generated in 8 weeks old FVB/n mice using the dermal punch method. Wounded or control skin fibroblasts were isolated by FACS as PDGFRα⁺CD45⁻EpCAM⁻ cells. g qRT-PCR analysis of the expression of selected genes in wound-derived fibroblasts as compared to their expression in fibroblasts from normal skin. n = pool of 8 mice per group. Data are presented as mean ± s.d. of biological repeats; Welch’s t-test. Representative of three independent biological experiments. Source data are provided as a Source Data file