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Analysis of GPI-anchored proteins. Procyclic forms of the wild-type and the procyclin null mutant were labeled with [3H]ethanolamine (A) or [3H]myristic acid (B) and the delipidated (CMW-insoluble) fractions were extracted sequentially with 9% (vol/vol) butan-1-ol, 0.1% (wt/vol) Triton X-100, and 1% (wt/vol) SDS. Then 3 × 108 equivalents of [3H]ethanolamine-labeled cells and 108 equivalents of [3H]myristic acid-labeled cells were analyzed by SDS-PAGE and autoradiography.
