Figure 3

Parkin-mediated mitophagy facilitates the progression of NCP BVDV infection. A Western blot analysis of PINK1 and Parkin proteins in mock and NCP BVDV-infected cells (MOI = 5) at the indicated time points post infection. CCCP (10 μM) group was collected at 12 hpi. B, C IFA analysis of the recruitment of Parkin to damaged mitochondria (B) and co-localization of p62 with Parkin (C) in mock and NCP BVDV-infected (MOI = 5) cells at 48 hpi, as well as in CCCP treated cells (10 μM) at 12 hpi (Scale bar = 10 μm). D MDBK cells were transfected with siParkin or siNC for 24 h. Then, the cells were infected with NCP BVDV (MOI = 5), Western blot assays were performed to determine VDAC1 and MFN2 expression at 48 hpi. E Western blot analysis of VDAC1 expression in MDBK cells stably expressing Parkin following NCP BVDV infection at 48 hpi. F MDBK cells were transfected with siParkin or siNC for 24 h. Then, the cells were infected with NCP BVDV (MOI = 5), qRT-PCR (left) and Western blot (right) assays were performed to determine the viral replication and progeny at 24 hpi and 48 hpi. G MDBK cells stably expressing Parkin were infected with NCP BVDV (MOI = 5), Western blot (left) and qRT-PCR (right) assays were performed to determine the viral progeny and replication at 48 hpi. Data are given as means ± standard deviation (SD) from three independent experiments. P values were calculated using Student t test. An asterisk indicates a comparison with the indicated control. *P < 0.05; **P < 0.01; ns: not significant.