Fig 1: The decline in number of mitochondria in CSFV-infected cells was related to mitophagy(A) Changes of mitochondrial proteins in CSFV-infected PK-15 cells. PK-15 cells were mock-infected or infected with CSFV (MOI = 1), and whole cell lysates (WCL) were prepared at 24, 36 and 48 hpi. The expression of mitochondrial proteins, including MFN2, HSP60, VDAC1, TOM20 and COX4, were analyzed by Western blotting. CSFV infection was verified by immunoblotting with anti-CSFV Npro antibody. GAPDH was used as an internal loading control. The histograms on the right showed the statistical analysis of the protein band intensity (mean ± SD; n = 3; *P < 0.05; *P< 0.01; ***P < 0.001). P values were calculated using two-way ANOVA. (B) Changes of mitochondrial proteins in CSFV-infected 3D4/2 cells were analyzed as in (A). (C) Changes of mitochondrial proteins in CSFV-infected PK-15 and 3D4/2 cells treated with 3-MA. PK-15 and 3D4/2 cells infected with CSFV (MOI = 1) in the presence or absence of 3-MA (5 mM) at 48 hpi. Expression of mitochondrial matrix proteins including HSP60 and COX4 were by Western blotting. Inhibition of autophagy determined by the detection of LC3-II expression. CSFV infection was verified by immunoblotting with anti-CSFV Npro antibody. GAPDH was used as an internal loading control. The lower histograms showed the statistical analysis of the intensity of mitochondrial protein bands (mean ± SD; n = 3; *P< 0.01; ***P < 0.001). P values were calculated by two-way ANOVA. (D) Changes of mitochondrial proteins in CSFV-infected PK-15 and 3D4/2 cells treated with BafA1. PK-15 and 3D4/2 cells were infected with CSFV (MOI = 1) in the presence or absence of BafA1 (5 nM) at 48 hpi. HSP60 and COX4 were analyzed as in (C). Inhibition of the autophagy flux was evaluated by detecting LC3-II and SQSTM1. The lower histograms showed the statistical analysis of the intensity of mitochondrial protein bands (mean ± SD; n = 3; *P< 0.5; ***P < 0.001). P values were calculated by two-way ANOVA.