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. 2020 Jan 17;9(1):234. doi: 10.3390/cells9010234

Figure 6.

Figure 6

Effects of l-cysteine on MGO-induced cell toxicity and ROS generation in HEK 293 cells. (A) Cell viability of HEK 293 cells with MGO (500 μM) and various concentrations of l-cysteine (0.1, 0.5, and 1.0 mM) and analyzed using the MTT assay. (B) MGO-induced LDH production was evaluated by LDH assay in HEK 293 cells. (C) Representative cytograms of Annexin V-FITC and PI staining of MGO-induced HEK 293 cells. Cells were pre-treated with several concentrations l-cysteine for 1 h, then incubated with MGO (500 μM) for 24 h. After 24 h, the concentrations of viable (Annexin V-FITC and PI negative cells), early-stage apoptotic (Annexin V-FITC positive, PI negative cells), late-stage apoptotic (Annexin V-FITC positive, PI-positive cells), and necrotic (PI-positive cells) cells were analyzed by flow cytometry. (a) control; (b) 500 μM MGO; (c) MGO+l-cysteine (0.1 mM); (d) MGO+l-cysteine (0.5 mM); (e) MGO+l-cysteine (1.0 mM); (f) MGO+AG (1.0 mM) as a positive control. (D,E) Percentage of control, early-stage apoptotic, late-stage apoptotic, and necrotic cells as analyzed by flow cytometry. (F) MES13 cells were pretreated with l-cysteine for 1 h, followed by 500 μM MGO for 1 h. Green fluorescence (ROS generation) from DCF-DA was examined by FAC analysis. Quantitative measurements of fluorescent intensity were evaluated using an FAC analysis system. (a) control vs 500 μM MGO; (b) 500 μM MGO+ l-cysteine (0.1 mM); (c) MGO+l-cysteine (0.5 mM); (d) MGO+l-cysteine (1.0 mM); (e) MGO+ AG (1.0 mM) as a positive control. All data are presented as mean ± SEM. n = 3 (### p < 0.001 vs. Control, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. MGO 500 μM).