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

Figure 1

The effects of BoHV-1 infection on the expression of Nrf2-regulated downstream targets. (a, c, d, and e) MDBK cells in 60 mm dishes were mock infected or infected with BoHV-1 at an MOI of 0.1 for 4, 8, 16, 24, 36, and 48 h. The cell lysates were then prepared for Western blots to detect the expression of Nrf2, HO-1, NQO1, and VP16 using the Nrf2 antibody (Abcam, cat# ab137550, 1 : 500), HO-1 monoclonal antibody (mouse) (Enzo Life Sciences, cat# ADI-OSA-110-D, 1 : 1000), NQO1 polyclonal antibody (ABclonal, cat# A0047, 1 : 1000), and VP16 antibody (a gift from Prof. Vikram Misra at the University of Saskatchewan, 1 : 2000). (b) MDBK cells were seeded into 24-well plates. After overnight incubation, the cells were infected with BoHV-1 at an MOI of 0.1. After infection for 4, 8, 16, 24, 36, and 48 h, the cells were collected and cell numbers were counted using a Trypan-blue exclusion test. Data shown are representative of three independent experiments. (f) The band intensity was analyzed with software ImageJ. Each analysis was compared with that of uninfected control at each time point, which was arbitrarily set as 100%. These images are representative of those from three independent experiments. (g) Total RNA was prepared at 16 and 24 hpi in MDBK cells, and the mRNA levels of Nrf2, HO-1, and NQO1 were measured by qRT-PCR. Each analysis was compared with that of uninfected control, which was arbitrarily set as 100%. Data represent three independent experiments. The significance was assessed with Student’s -test ().

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