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. 2002 Mar 4;195(5):593–602. doi: 10.1084/jem.20011500

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Copyright © 2002, The Rockefeller University Press

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Figure 3. — Localization of filovirus proteins in lipid rafts in infected cells. (A) Primary human monocytes were infected with MBGV. After 24 h cells were lysed in 0.5% Triton X100 and detergent-soluble (S) and -insoluble (I) fractions were separated by centrifugation, samples were irradiated (2 × 10⁶ R), and analyzed by immunoblotting with a guinea pig anti-MBGV antibody to detect viral proteins NP and VP35/VP40 (lanes 3 and 4); lanes 1 and 2, uninfected control; lane 5, inactivated MBGV (1 μg). NS, nonspecific band. (B) HepG2 hepatocytes were infected with EBOV-Zaire, lysed, irradiated (6 × 10⁶ R), and rafts (R) and soluble (S) fractions were prepared by ultracentrifugation 24 h after infection. Ebola GP and VP40 were detected by immunoblotting. (C) Ebola-infected Vero E6 cells were irradiated (4 × 10⁶ R), fixed and stained for Ebola virus (red) and GM1 (green) at 4°C and imaged by confocal microscopy. (Left panel) single section; (right panel), 3-D reconstruction of the compiled data.

Figure 3. — Localization of filovirus proteins in lipid rafts in infected cells. (A) Primary human monocytes were infected with MBGV. After 24 h cells were lysed in 0.5% Triton X100 and detergent-soluble (S) and -insoluble (I) fractions were separated by centrifugation, samples were irradiated (2 × 10⁶ R), and analyzed by immunoblotting with a guinea pig anti-MBGV antibody to detect viral proteins NP and VP35/VP40 (lanes 3 and 4); lanes 1 and 2, uninfected control; lane 5, inactivated MBGV (1 μg). NS, nonspecific band. (B) HepG2 hepatocytes were infected with EBOV-Zaire, lysed, irradiated (6 × 10⁶ R), and rafts (R) and soluble (S) fractions were prepared by ultracentrifugation 24 h after infection. Ebola GP and VP40 were detected by immunoblotting. (C) Ebola-infected Vero E6 cells were irradiated (4 × 10⁶ R), fixed and stained for Ebola virus (red) and GM1 (green) at 4°C and imaged by confocal microscopy. (Left panel) single section; (right panel), 3-D reconstruction of the compiled data.