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Fig. 4. Low- and intermediate-density fractions originate from poorly solubilized microdomains of the plasma membrane. (A) 2x10⁷ Jurkat cells were biotin-labeled and lysed at 4°C in MNE supplemented with 0.5% Triton X-100. After overnight centrifugation in a three-step (42.5 %, 30%, 5%) sucrose gradient, 12 fractions of 280 ml were collected from the top and four sequential fractions were pooled (1-4, 5-8, 9-12). An equal amount of each pool was either left untreated (cell lysate) or was immunoprecipitated with avidin coated beads (IPP: Avidin) before SDS-PAGE analysis and blotting with avidin (left) or anti-CD9 antibody (right). (B) 2x10⁷ cells were lysed and fractionated by sucrose gradient as above. A pool of fractions 2-3 (F2-3, left) or 6-7 (F6-7, right) were washed, concentrated and analysed by electron microscopy. Bar, 1 µm.

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