Figure 7
Inhibition of Gpx activity induces T-cell hyporesponsiveness. (a) The splenocytes isolated from spleen of WT mice with NAC (5 mM, pharmacological antioxidant) or without, and pretreated MS (0.4 mM) for 48 h, then treated Con A (5 μg/ml) for 8 h. Then, we performed cytokine assay. (b) Con A-induced mRNA expressions of Th1 cytokines such as IL-2, IFN-γ and TNF-α in MS (0.4 mM, Gpx inhibitor)-pretreated Jurkat T cell with NAC (5 mM, pharmacological antioxidant) or without. Values are expressed as the mean±S.E.M. of three different experiments conducted in triplicates. *P<0.05, control versus Con A, ##P<0.05, Con A versus pretreatment of MS then Con A, #P<0.05, pretreatment of MS then Con A versus pretreatment of MS in the presence of NAC then Con A. (c) BrdU incorporation assay of Con A-induced cell proliferation in MS (0.4 mM, Gpx inhibitor)-pretreated Jurkat T cell with NAC (5 mM, pharmacological antioxidant) or without using the Cell Signaling BrdU Cell Proliferation Assay Kit. Values are expressed as the mean±S.E.M. of three different experiments conducted in triplicates. *P<0.05, control versus Con A, ##P<0.05, Con A versus pretreatment of MS then Con A, #P<0.05, pretreatment of MS then Con A versus pretreatment of MS in the presence of NAC then Con A. (d) Immunoblots of PLCγ and IκB phosphorylation in Con A-induced cell proliferation in MS (0.4 mM, Gpx inhibitor)-pretreated Jurkat T cell with NAC (5 mM, pharmacological antioxidant) or without
