Fig. 4.

SSc exosome-stimulated macrophages activate SSc fibroblasts

Monocyte-derived human macrophages were plated in the bottom chamber of Transwell plates, and were differentiated and activated with 2 × 10⁹ healthy (HC exo) or SSc patient fibroblast-derived exosomes (SSc exo) for 48 h. At conclusion of activation, Transwells containing SSc patient fibroblasts were inserted into wells. (A) Schematic diagram of culture conditions. (B–F) Total RNA was extracted from mono- (HC/SSc exo-mono) or co-cultured (HC/SSc exo-Co) SSc-fibroblasts. Relative mRNA levels were measured using Taqman real-time PCR. *P < 0.05, **P < 0.01, ***P < 0.0005, ****P < 0.0001; data shown are representative of results obtained from analysis of 4 healthy donors, 4 SSc patients. Two technical replicates were analysed in three separate experiments. Error bars represent s.d.