Assembly and release of infectious hepatitis C virus involving unusual organization of the secretory pathway

Figure 5 Calnexin and N-linked glycosylation are involved in the release of hepatitis C virus particles via a non-classical secretion path in baby hamster kidney-West Nile virus cells. A: BHK-21 and BHK-WNV cells were transfected with the HCV expression plasmid system. IF was performed three days later with anti-ERGIC-53 (green) and anti-CRT (red) antibodies, followed by confocal microscopy analysis; B: Same protocol as in (A) with BHK-WNV cells transfected with the HCV-coding plasmids; C: Twelve hours after transfection with the HCV expression plasmid mix, parental BHK cells or BHK-WNV cells were treated, or not, with N- (DNJ) or O- (PAG, ALL) glycosylation inhibitors; materials released in SN (top and middle panels) or cell lysates (bottom panel) were collected, incubated with or without Endo-H and analyzed by Western blot. HCV: Hepatitis C virus; BHK-WNV: Baby hamster kidney-West Nile virus; ERGIC-53: Endoplasmic reticulum-Golgi intermediate compartment-protein of 53 kDa; CRT: Calreticulin; SN: Supernatants; DNJ: Deoxynojirimycin; Endo-H: Endo-β-N-acetylglucosaminidase H.