Figure 2

Baseline antioxidant levels in sorted CD4⁺ T-cell subpopulations reflect their susceptibility to AF. (a) Total GSH levels in sorted CD4⁺ T-cell subpopulations. CD4⁺ T-cell subpopulations were separated by sorting and baseline GSH levels were measured. Data points represent the means from three independent experiments normalized to T_N cells. Data are shown as median±interquartile range (n=3; P<0.001; repeated measures ANOVA). (b) Total thiol content in sorted CD4⁺ T-cell subpopulations. The thiol content was measured, in all CD4⁺ T-cell subpopulations, by FACS after labeling cells with ThiolTracker violet dye. The data displayed in both the dot plot and the x, y graph are expressed as the median of fluorescence intensity. On the ‘x’ axis of the x, y graph, the level of CD4⁺ T-cell differentiation was associated, for clarity purposes, to an increasing score ranging from 0 to 3 (0=T_N; 1=T_CM; 2=T_TM; 3=T_EM; P<0.05; Spearman’s correlation test). (c) Replication of the effects of AF on CD4⁺ T cells by the TrxR inhibitor arsenic trioxide (As₂O₃). The dot plots show the effect of AF and As₂O₃ on total CD4⁺ T cells after 48 h of treatment, analyzed by CD27 (y axis) and Annexin V (x axis) staining. The dot plot shows data from one representative experiment out of three with similar results