Figure 1
NCP BVDV infection induces the formation of mitophagosomes. A Western blot analysis of NS4B protein in mock and NCP BVDV-infected (MOI = 1, 5, 10) cells at 48 hpi (upper) or at 0 hpi, 24 hpi, 48 hpi and 72 hpi (MOI = 5) (under). Equal amounts of protein from mock and NCP BVDV-infected cells were separated using SDS-PAGE and transferred to PVDF membranes. The membranes were probed with NS4B antibody. B RNA from NCP BVDV-infected (MOI = 5) cells was collected, and BVDV Npro gene was measured by qRT-PCR. C NCP BVDV-infected (MOI = 1, 5, 10) cells were bound with BVDV NS4B antibody and then labelled with fluorophore-conjugated antibody or matched isotype controls and analyzed by flow cytometry at 48 hpi. D IFA analysis of NS4B expression in cells infected with NCP BVDV (MOI = 5) at 48 hpi (Scale bar = 50 μm). E Western blot analysis of LC3-II, p62, VDAC1 and MFN2 proteins in mock and NCP BVDV-infected cells at 24 hpi and 48 hpi. F, G IFA analysis of the recruitment of LC3-II to damaged mitochondria (F) and the recruitment of p62 to damaged mitochondria (G) in mock and NCP BVDV-infected (MOI = 5) cells at 48 hpi. (Scale bar = 10 μm). 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; ***P < 0.001; ns: not significant.