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. 2017 Sep 1;8:988. doi: 10.3389/fimmu.2017.00988

Figure 3.

Figure 3

HOCl- or H2O2-induced serum AOPPs increased nitric oxide (NO) and reactive oxygen species (ROS) production in MSCs. (A) Quantification of NO production in MSCs in culture with human serum AB (SAB) or oxidized SAB (SAB400, SAB1000, and SABH2O2; n = 7 for each condition) or SSc patient serum at 24 h. Sera from patient (PS) were divided in two groups depending on AOPP levels: <400 μmol/L (PS<400; n = 9) or >400 μmol/L (PS>400; n = 7) or pooled in a single group (PSpool; n = 16). NO production was measured using 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate probes (Molecular Probes, ThermoFisher Scientific). (B) Quantification of ROS production in MSCs in same conditions as in (A). ROS production was measured using 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate probes (Molecular Probes, ThermoFisher Scientific). (C) Gene expression fold change of Sod2 antioxidant marker in same conditions as in (A) (PS<400 and PS>400, n = 4; PSpool, n = 8). Data were normalized to 1 for MSCs in SAB-containing medium. *p < 0.05 versus SAB at same time point.