Fig. 4

ROS activity in response to TCDD in WT and Jdp2^−/− MEFs. A MEFs were incubated with 10 nM TCDD for 2 h, stained with 0.25 M CM-H₂-DCFDA, and examined by flow cytometry, as described in the “Methods” section. B ROS activity was measured in WT and Jdp2^−/− MEFs in response to exposure to 10 nM TCDD for 0, 2, 6, or 24 h. ROS production was detected using CM-H₂DCFDA, as described in the “Methods” section. Representative fluorescence images of ROS generation in WT (top) and Jdp2^−/− (bottom) MEFs are shown. C The data obtained in the fluorescence images of ROS levels detected using CM-H₂DCFDA after treatment with TCDD were analyzed using ImageJ software. The fluorescence intensity of WT MEFs and Jdp2^−/− MEFs after TCDD exposure was set at 1.0. Values represent the mean ± SEM (n = 5). Data were analyzed using two-way ANOVA with the Bonferroni post hoc test (*p < 0.05). D–H Jdp2 controls ROS production and antioxidation reaction in MEFs. D Levels of 8-oxo-dGuo in WT and Jdp2^−/− MEFs in the presence or absence of TCDD (10 nM) for 2 h in cells harvested at 24 h. E Levels of malondialdehyde (MDA) in in WT and Jdp2^−/− MEFs in the presence or absence of TCDD (10 nM) for 2 h in cells harvested at 24 h. F, G Levels of total glutathione (GSH) (F) and the GSH/oxidized glutathione (GSSG) ratio (G) in WT and Jdp2^−/− MEFs with or without exposure to TCDD (10 nM) for 2 h in cells harvested at 24 h. H Relative NQO1 enzyme activity in WT and Jdp2.^−/− MEFs with or without exposure to TCDD (10 nM) for 2 h in cells harvested at 24 h. Data are presented as the mean ± SEM (n = 3). Data were analyzed using one-way ANOVA with the Turkey post hoc test (*p < 0.05 and **p < 0.01)