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. 2022 Dec 1;23(23):15137. doi: 10.3390/ijms232315137

Figure 2.

Figure 2

Antioxidant activities of NAst. (a) Proliferation of SHSY-5Y cells supplemented with 3.7 ng/µL of NAst and synthetic astaxanthin. Cells were counted in triplicate by the Trypan blue exclusion method. Data are reported as cell numbers ± SDs. (b) Effects of NAst on intracellular ROS levels in SHSY-5Y cells. Cells were supplemented with different amounts of NAst for 24 h. Intracellular ROS levels were measured in live cells using the probe DCFH-DA by means of spectrofluorimetric analysis. Data are reported as percentages in comparison to controls ± SEMs. (c) Total glutathione evaluation in SHSY-5Y cells supplemented with 3.7 ng/μL NAst by luminescent assay. Data are shown as the means ± SDs of at least three independent experiments. ** p ≤ 0.01. (d) NQO1 activity determination in lysates from SHSY-5Y cells supplemented with NAst for 24 h. Dicoumarol was used to specifically inhibit the enzyme. Data are reported as fold increases in comparison with untreated cells ± SDs; n = 4. ** p ≤ 0.01. (e) Representative micrographs of SHSY-5Y cells stained with the lipid peroxidation sensor BODIPY C11. Cells were supplemented with 3.7 ng/µL of NAst for 24 h. Oxidative stress was induced using 130 µM of TBH for 30 min. Scale bars: 10 µm. (f) Quantitative analysis of the red/green BODIPY C11 fluorescence ratio performed using ImageJ software. Five randomly chosen fields were analyzed for each condition. Error bars indicate SDs. ** p ≤ 0.01, *** p ≤ 0.001.