The Differential Expression of Mitochondrial Function-Associated Proteins and Antioxidant Enzymes during Bovine Herpesvirus 1 Infection: A Potential Mechanism for Virus Infection-Induced Oxidative Mitochondrial Dysfunction

Figure 4

The effects of DPI treatment on the expression of NRF1, NRF2, and TFAM following BoHV-1 infection. (a) MDBK cells in 24-well plates pretreated with DPI (5 μM) or DMSO control for 1 h were infected with BoHV-1 () along with corresponding chemical treatment. After infection for 16 h, cellular ROS levels were determined using H2DCFDA (5 μM, 30 min) (Sigma-Aldrich, St. Louis, MO, USA) and quantitatively analyzed using software Image-pro Plus 6. Data shown are means of three independent experiments. Significant differences (), as determined by the Student -test. (b) MDBK cells in 24-well plates were treated with DPI or MDSO control and infected by BoHV-1 () at the same condition as described in (a). Finally, the cell cultures were subjected to frozen-thawing twice, and viral production was determined using MDBK cells with results expressed as TCID50/mL. (c–e) MDBK cells in 60 mm dishes pretreated with DPI (5 μM) or DMSO control for 1 h were infected with BoHV-1 () in the presence of DPI or DMSO control for 16 h; the cell lysates were prepared for Western blots to detect the expression of NRF1 (c), NRF2 (d), and TFAM (e). Data shown are representative of three independent experiments. (f) The relative band intensity was analyzed with software ImageJ, and each analysis was compared with that of the uninfected control which was arbitrarily set as 100%. Data are means of three independent experiments. Significance was assessed with the Student -test ().

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