Fig. 3.

SiRNA CCDC6 prevents M1 virus-induced expression of antiviral indicators and activates replication of M1 virus. (a) Pretreatment with CCDC6 siRNA 24 h after UM-UC-3 Huh-7 and HCT116 cells are infected with M1 virus (MOI = 1). Phase contrast microscope image of 48 h. Scale bar: 100 μm. (b and c) M1-GFP virus (MOI = 1) was infected for 24 h, the M1 virus infection rate was measured by flow cytometry (b), and the comparison of the GFP positive proportion of total cell number between CCDC6 siRNA group and negative control (NC) siRNA group for UM-UC-3, Huh-7 and HCT116 cells (c). (d) After 48 h of M1 virus infection, the M1 virus titer (MOI = 1) was measured by the TCID50 assay. (e) Expression of E1 mRNA of the M1 virus was performed by qRT-PCR 4 h after infection with M1 virus (MOI = 10). Relative gene manifestation levels were standardized to β-actin. (f) M1 viral protein levels were distinguished by Western blot 24 h after M1 virus infection (MOI = 1). (g) After 12 h of treatment with M1 virus (MOI = 10), mRNA levels of IRF7, IFIT1, IRNB, IFIH1, and IRF3 were detected by qRT-PCR (mean ± SD). The expression of genes was normalized to β-actin. The data are shown in mean ± SEM. * P < 0.05; ** P < 0.01; ***P < 0.001; N.S., not significant. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.