Bovine Herpesvirus 1 Productive Infection Led to Inactivation of Nrf2 Signaling through Diverse Approaches

Figure 2

The effects of Trolox on the expression of Nrf2 and its downstream targets. (a, b, and c) MDBK cells in 60 mm dishes pretreated with Trolox (1 mM) or DMSO control for 1 h were infected with BoHV-1 (); in the presence of Trolox or DMSO control for 24 h, the cell lysates were prepared for Western blots to detect the expression of Nrf2 (a), HO-1 (b), and NQO1 (c). (e, f, and g) MDBK cells in 60 mm dishes pretreated with Trolox (1 mM) or DMSO control for 1 h were exposed to tBHP in the presence of Trolox or DMSO control for 2 h; the cell lysates were prepared for Western blots to detect the expression of Nrf2 (e), HO-1 (f), and NQO1 (g). (d and h) The relative band intensity was analyzed with software ImageJ, and each analysis was compared with that of uninfected control at each time point, which was arbitrarily set as 100%. Data shown are representative of three independent experiments. (i) MDBK cells in 24-well plates pretreated with Trolox at indicated concentrations or MDSO control were infected with BoHV-1 () for 24 hours in the presence of an inhibitor or DMSO. The cell cultures were subjected to frozen-thawing twice, and viral yield was determined with the results being expressed as TCID₅₀/mL. (j) The cytotoxicity of Trolox in MDBK cells for 24 h was analyzed by Trypan-blue exclusion. The significance was assessed with Student’s -test ().

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