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S3). Careful analysis of the data indicated that the virion particles collected at early stage after infection were more sensitive to the neutralizing antibody than those collected at late stage after infection. The data suggest that viruses produced at early stage after infection rely on the apoE-dependent infection process more than those produced at late stage after infection thus show higher infectivity. The data also suggest that the infectious virion particles collected at the late stage of infection also contain some apoE proteins, and that the late stage of infections are, at least in part, facilitated by apoE. These results indicate that the secretion of apoE is likely related with the production of infectious [http://en.wikipedia.org/wiki/Parvulin Parvulin] virion particles but not with the secretion of HCV RNA-containing particles and that the composition of virion particles may change during HCV infection cycle. Previous studies had revealed that the HCV RNA-containing particles produced in cell culture display a broad range of buoyant densities, suggesting the existence of HCV virions with different components (Hijikata et al., 1993?and?Pumeechockchai et al., 2002). The authors used mixtures of HCV particles produced at various stages of infection. To investigate the properties of HCV particles produced at specific [http://www.selleckchem.com/products/ve-821.html check details] stages of infection, we harvested the culture media of HCV-infected cells every 12?h and analyzed the samples by isopycnic sucrose density gradient. The amounts of HCV RNAs in each fraction of sucrose gradient were quantified by RT-qPCR, the titers of infectious HCV particles in each fraction were determined by infection of na?ve Huh7.5.1 cells (Fig. 4). The majority of viral RNAs were fractionated at buoyant densities between 1.15 and 1.17?g/ml throughout the infection course (black lines in Fig. 4A�CC). Although the high buoyant density fractions contained the highest amounts [http://www.selleckchem.com/products/BEZ235.html BEZ235 mouse] of viral RNA, only low levels of infectious viruses were found in these fractions. This indicates that most of the viral RNAs in the high density fractions are not infectious. The fraction containing the highest titer of infectious HCV was the lowest buoyant density fraction (