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. 2020 Oct 23;11:532177. doi: 10.3389/fimmu.2020.532177

Figure 4.

Figure 4

Effects of patients’ according to clinical features: disease duration, pulmonary fibrosis and methotrexate intake. Levels of Col1A1 mRNA, Col1A2, MMP1, and CCL2 were assessed in fibroblasts stimulated with 1,000 MPs/µl of HD (n=12) or SSc (n=12) by reverse transcriptase quantitative polymerase chain reaction (RT qPCR) and expressed as a Col1a1/MMP1 (A, D, G) or Col1a2/MMP1 ratio (B, E, H), or in relation to ΔΔCt for housekeeping gene and for DMEM 10% FBS as the baseline condition for CCL2 (C, F, I). Then, for each analysis, we compared the effect of MPs from HD or SSc patient according to different clinical parameters: duration between the onset of the first symptoms other than Raynaud’s phenomenon less than (n=8) or greater than (n=4) 3 years (A–C), presence (n=3) or absence (n=9) of lung fibrosis (D–F) and patients treated (n=6) or not treated (n=6) with methotrexate (G–I). The dashed line represents the fibroblast condition treated only with DMEM 10% FBS; Box-plots represent the extreme values, the first and third quartiles and the medians. **p < 0,01 (A–C, E, F, H, I), *p < 0.05 (A, C, D, F–I) by Mann-Whitney test. MPs, microparticles; HD, healthy donors; SSc, systemic sclerosis; Col1a1, Type 1 Collagen a1; Col1a2, Type 1 Collagen a2; MMP1, Matrix Metallopeptidase 1; CCL2, C-C Motif Chemokine Ligand 2; MTX, methotrexate; FBS, fetal bovine serum.