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. 2004 Nov;78(21):12047–12053. doi: 10.1128/JVI.78.21.12047-12053.2004

FIG. 1.

FIG. 1.

HCV core protein associates with a detergent-resistant membrane biochemically similar to classical lipid rafts. (A) FLRP1 cells (Huh7 cells harboring HCV replicons) were lysed, and aliquots were incubated at 4°C in the absence or presence of 1% Triton X-100 (Tx-100). The lysates were overlaid with a 5 to 40% OptiPrep gradient. Following ultracentrifugation, fractions were collected from the top of the tube and the proteins were precipitated and analyzed by Western blotting using an antibody against HCV core protein. The locations of fractions containing membranes, DRMs, and soluble proteins are indicated. (B) FLRP1 and Huh7 cells were lysed in cold 1% Triton X-100 and loaded in an 8 to 35% Nycodenz gradient. Following ultracentrifugation, fractions were collected from the top of the tube and the proteins were precipitated and analyzed by Western blotting using antibodies against core protein, caveolin-1, aerolysin (to detect GPI-linked proteins), and calnexin. (C) FLRP1 cells were treated with 10 mM mβCD or αCD for 30 min prior to lysis and analyzed as described above for HCV core protein. Memb., membrane.