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

Figure 2.

Figure 2

HOCl- or H2O2-induced serum AOPPs and systemic sclerosis (SSc) patient serum increased the number of senescent MSCs. (A) quantification of SA-β-gal activity in senescent MSCs in culture with human serum AB (SAB) or oxidized SAB (SAB400, SAB1000, and SABH2O2) or SSc patient serum at different time points: day 3, 6, or 10 (n = 8). Senescence was measured using the quantitative cellular senescence assay kit (Cells Biolabs) and expressed as relative fluorescence unit (RFU). Sera from patient (PS) were divided in two groups depending on AOPP levels: <400 μmol/L (PS<400; n = 11) or >400 μmol/L (PS>400; n = 9) or pooled in a single group (PSpool; n = 20). Data were normalized to 100 for senescent MSCs detected in SAB-containing medium. (B) Representative photographs of SA-β-gal staining of MSCs cultured in same conditions as in (A), at day 6. (C) Gene expression fold change of different senescence markers: p16, p21, and p27 in same conditions as in (A) (PS<400 and PS>400, n = 4; PSpool, n = 8). Data were normalized to 1 for apoptotic MSCs detected in SAB-containing medium. *p < 0.05 versus SAB at same time point.