FIGURE 2:
PrPC redistribution in continuous sucrose density gradient fractions following CD95/Fas treatment. CEM cells, either untreated or treated with anti–CD95/Fas (250 ng/ml for 30 min at 37°C), were lysed in lysis buffer, and postnuclear supernatant was prepared and loaded on top of a continuous 10–40% (wt/vol) sucrose gradient. After centrifugation, fractions were collected, separated by SDS–PAGE, and analyzed by Western blot. Fractions obtained after sucrose density gradient centrifugation, either untreated or treated with anti–CD95/Fas (250 ng/ml) cells, were analyzed using anti-PrP mAb (SAF 32). As controls, fractions obtained after sucrose density gradient centrifugation from untreated cells were analyzed using a polyclonal anti–VDAC-1 or a monoclonal anti-CD71.