Fig 2: MnSOD in T-lymphocytes is efficiently removed using an inducible knock-out strategy. Tamoxifen was administered to MnSOD knock-out (Cre+/-) and control (Cre-/-) mice for five consecutive days followed by two weeks to allow full recombination and loss of MnSOD. Splenic T-lymphocytes were isolated from mice and analyzed in either their naïve or activated (cultured with anti-CD3 and anti-CD28 beads for 96 h) states. A. Left, MnSOD gene and right, CuZnSOD gene mRNA levels assessed by quantitative real-time RT-PCR. B. Left, representative western blot of MnSOD and CuZnSOD. Right, quantification of respective proteins. C. Left, representative activity gel of MnSOD and CuZnSOD. Right, quantification of respective proteins. #p < 0.01 versus naïve Cre-/-; *p < 0.01 versus activated Cre-/- by parametric Student's t-test or nonparametric Mann-Whitney U test where appropriate.

Fig 3: mRNA and protein expression levels of Cu/Zn-SOD, Mn-SOD, and CAT in the small intestine tissue of mice. Values presented are the mean ± standard deviation (N = 10/group). a–eMean values with different letters in the same bars are significantly different (p < 0.05) according to Duncan's multiple range test. LB-FSM group: mice treated with Lactobacillus bulgaricus-fermented soybean milk; LP-CQPC01-FSM group: mice treated with Lactobacillus plantarum CQPC01-fermented soybean milk; U-FSM group: mice treated with unfermented soybean milk

Fig 4: Advanced glycation end products (AGEs) impaired the viability of bone marrow mesenchymal stromal cells (bMSCs) through increased reactive oxygen species (ROS) production attributable to selective reduction of superoxide dismutase (SOD)-1 and SOD-3 expression. A through C, Culture of bMSCs with AGE-BSA (with concentration from 100 to 400 µg/mL with BSA as control) significantly increased intracellular ROS (A) and early (B) and late apoptosis (C) in a dose-dependent manner. n=6/group. *P<0.0001 for 0 vs 400 µg/mL AGE-BSA; # P<0.0001 for 400 µg/mL BSA vs 400 µg/mL AGE-BSA; + P<0.001 for 0 vs 200 µg/mL AGE-BSA. D through H, The representative images of flow cytometric analysis (D and E) and quantification of intracellular ROS (F) and early (G) and late apoptosis (H). AGE-BSA treatment (400 µg/mL with BSA as control) significantly increased intracellular ROS, and apoptosis of bMSCs. Inhibition of ROS production with N-acetylcysteine (NAC) significantly prevented AGE-induced increase in intracellular ROS, and apoptosis of bMSCs (n=6/group; *P<0.01 for WT+AGE vs WT+BSA; + P<0.01 for WT+AGE+NAC vs WT+AGE). I through L, The representative images (I and K) and quantification (J and L) of Western blot analysis. AGE-BSA treatment (400 µg/mL) significantly decreased the protein levels of SOD-1 and SOD-3 in bMSCs (n=3; *P<0.05 for WT+AGE vs WT+BSA; J). The expression levels of SOD-1, SOD-3, and glutathione peroxidase-1 (Gpx-1) were significantly increased in bMSCs from triple-transgenic (TG) mice compared with bMSCs from wild-type (WT) mice (n=3; *P<0.05 for TG vs WT; L). To determine if decreased levels of SOD-1 and SOD-3 contributed to AGE-induced increase of ROS production in bMSCs and subsequent increase in apoptosis, experiments were repeated with bMSCs from mice with concomitant overexpression of SOD-1, SOD-3, and Gpx-1 (TG-bMSCs). Indeed, AGE-induced ROS and apoptosis were significantly decreased in TG-bMSCs compared with WT-bMSCs (n=6/group; # P<0.05 for TG+AGE vs WT+AGE). Statistical differences were determined with 1-way ANOVA with Bonferroni post hoc test or Kruskal-Wallis test with Dunn post hoc test. All data were shown as means±SD. FITC indicates fluorescein isothiocyanate; DCF, dichlorofluorescein; PI, propidium iodide; SSC-A, side scatter area.

Fig 5: Concomitant overexpression of triple antioxidant enzymes effectively preserved the muscle fibres of the ischemic limb. The muscle fibres reduced in size and became largely disconnected in the WT control mice with limb ischaemia. CD31 staining showed that capillary density was low in the limb muscle after ischaemia for 3, 7 and 21 days. On the other hand, the muscle fibres appeared to remain long and large, as well as well-connected in the limb muscles at day 21 after ischaemia in the TG mice with concomitant global overexpression of SOD1, SOD3 and Gpx-1. CD31 staining demonstrated that the capillary density remained largely intact (similar to the non-ischemic limb). Normal: normal right limb; ischaemia; ischemic left limb. Red arrow: capillary vessels. BL: before ligation. Scale bar: 50µm