FIG 5.

Effect of degradation of SM by sphingomyelinase (SMase) on the morphology of DMVs. JFH1/SGR cell lysates were treated with 1% Triton X-100 for 60 min on ice and fractionated by discontinuous sucrose gradient centrifugation. (A) Equal volumes of the recovered concentrated fractions were analyzed by WB using antibodies against NS5A, calnexin, or caveolin 2. Fractions are numbered from 1 to 9 in order from top to bottom. (B) The DRM fraction (fraction 1) from the experiment shown in panel A was subjected to SMase treatment (10 U/ml) and examined using TEM. Representative DMVs and deformed vesicle membrane structures are shown. (C) In the control or SMase-treated fractions, the percentages and diameters of DMVs (left) and deformed (Def.) vesicles (right) were determined for 200 membrane structures in each and plotted as a percentage of the total number of vesicles.