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. 2018 Jun 5;15(6):1173. doi: 10.3390/ijerph15061173

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Effects of EESS on H₂O₂-induced ROS generation, DNA damage, and apoptosis in SW1353 chondrocytes. The cells were pretreated for 1 h with 400 µg/mL EESS, and then treated for 24 h with or without 1 mM H₂O₂ in the absence or presence of 10 µM ZnPP. (A) After incubation with 10 µM JC-1 for 20 min, JC-1 fluorescence intensity was detected by flow cytometry. Each point represents the mean of two independent experiments; (B) The comet assay was performed to detect cellular DNA damage, and representative photographs of the comets were taken by fluorescence microscopy (original magnification, 200×); (C) To quantify the degree of apoptosis, the cells were stained with FITC-conjugated Annexin-V and PI for flow cytometry analysis. The data are represented as the means ± SD obtained from three independent experiments (* p < 0.05 compared with the control group; ^# p < 0.05 compared with the H₂O₂-treated group; ^$ p < 0.05 compared with H₂O₂ and the EESS-treated group).

Effects of EESS on H₂O₂-induced ROS generation, DNA damage, and apoptosis in SW1353 chondrocytes. The cells were pretreated for 1 h with 400 µg/mL EESS, and then treated for 24 h with or without 1 mM H₂O₂ in the absence or presence of 10 µM ZnPP. (A) After incubation with 10 µM JC-1 for 20 min, JC-1 fluorescence intensity was detected by flow cytometry. Each point represents the mean of two independent experiments; (B) The comet assay was performed to detect cellular DNA damage, and representative photographs of the comets were taken by fluorescence microscopy (original magnification, 200×); (C) To quantify the degree of apoptosis, the cells were stained with FITC-conjugated Annexin-V and PI for flow cytometry analysis. The data are represented as the means ± SD obtained from three independent experiments (* p < 0.05 compared with the control group; ^# p < 0.05 compared with the H₂O₂-treated group; ^$ p < 0.05 compared with H₂O₂ and the EESS-treated group).