Fig 2: (A) Effect of SHP-1 on intracellular ROS. Cells were treated with SHP-1 (10, 20, 50, and 100 µM) for 2 hr. Then 0.75 M ethanol was added and incubated for 24 hr. DCFH-DA was added and reacted for 20 min. Cells were washed with PBS three times and observed under a fluorescent inverted microscope. a, The blank group (HepG2/CYP2E1 cells without treatment); b, the control group (HepG2/CYP2E1 cells with ethanol treatment only); c–f, 10–100 µM SHP-1. (B) Effect of SHP-2 on intracellular ROS. a, the blank group; b, the control group; c–f, 10–100 µM SHP-2. (C) Effect of SHP-1 on SOD activity. Values are mean ± SD (n = 3). (D) Effect of SHP-2 on SOD activity. Values are mean ± SD (n = 3). (E) Effect of SHP-1 on GSH. Values are mean ± SD (n = 3). (F) Effect of SHP-2 on GSH. Values are mean ± SD (n = 3). (G) Effect of SHP-1 on GGT activity. Values are mean ± SD (n = 3). (H) Effect of SHP-2 on GGT activity. Values are mean ± SD (n = 3). (I) Effect of SHP-1 on the expressions of SOD, GSH, and GGT. Cells were treated with SHP-1 (20, 50, and 100 µM) for 2 hr. Then, 0.75 M ethanol was added and incubated for 24 hr. SOD, GSH, and GGT levels were detected by Western blot. GAPDH was used as internal control. (J) Effect of SHP-2 on the expressions of SOD, GSH, and GGT. ### p < .001 represented the control group vs. the blank group. *p < .05 and ***p < .001 vs. the control group

Fig 3: IAB relieves oxidative stress in mice with AD via the regulation of the Nrf2 pathway. Compared with vehicle-treated AD mice, 28-day administration of IAB reduced the levels of (A and D) ROS, and enhanced the levels of (B and E) SOD and (C and F) GSH-Px in the serum and whole brains of mice with AD. Data are expressed as mean ± standard error of the mean (n=6). (G) IAB regulated the expression of proteins associated apoptosis and oxidative stress in the brains of AD mice (n=6). Quantification data was normalized by GAPDH. The mean fold of band intensity compared with CTRL group was presented respectively. #P<0.05, ##P<0.01 and ###P<0.001 vs. CTRL mice. *P<0.05, **P<0.01 and ***P<0.001 vs. vehicle-treated AD mice. AD, Alzheimer's disease; Bcl-2, B-cell lymphoma-2; Bax, Bcl-2-associated X; Bcl-xL, Bcl-extra large; CAT catalase; CTRL, control; D-gal, D-galactose; GSH-Px, glutathione peroxidase; HO, heme oxygenase; IAB, isoastilbin; L-Glu, L-glutamic acid; Nrf2, NF-E2p45-related factor 2; ROS, reactive oxygen species; SOD, superoxide dismutase.

Fig 4: Maternal antioxidant enzyme activity and gene expression in the liver at 19dG. (A) SOD activity (U/mg protein); (B) SOD gene expression; (C) SOD inmunostained area (%); (D) Representative micrograph of SOD-1 (40×); (E) GPx activity (mU/mg protein); (F) GPx gene expression; (G) GPx 1/2 inmunostained area (%); (H) Representative micrograph of GPx 1/2 (40×). Values are mean ± SEM, (C: control, n = 8; Cres: control + resveratrol, n = 10; MO: maternal obesity, n = 7; MOres: maternal obesity + resveratrol, n = 7). Within the same group (C vs. Cres and MO vs. MOres) means labelled with different letters differ, p < 0.05; * different in comparison to the respective control (MO vs. C and MOres vs. Cres), p < 0.05. I = interaction between maternal diet and maternal resveratrol intervention, n.s. = not significant.

Fig 5: Human PBMCs from DS individuals show the induction of antioxidant responses to counteract OS. Slot blot analysis of total protein carbonylation. Panel (A) (left hand side): Slot blot of representative samples from healthy donors (HD) and Down syndrome (DS) groups. A triplicate of eight samples per group is showed. Panel (B) (right hand side): Densitometric analysis of total protein carbonylation in healthy donors (HD) and Down syndrome (DS) groups. (C) Representative Western blot showing SOD-1, PRDX6, and GELSOLIN in PBMCs from DS and HD. (D) Quantification of panel C showing levels of total GELSOLIN. (E) Quantification of panel C showing levels of PRDX6 (E) Quantification of panel C showing levels of total GELSOLIN. (F) Quantification of panel (C) showing levels of SOD1. Densitometric values shown in the bar graph are the mean of six samples for each group normalized for total load and are given as percentage of HD, set as 100%. Statistical significance was determined using Student t-test analysis (* p < 0.05, ** p < 0.01, *** p < 0.001).