Figure 3.

Depletion of Gpi11p results in a block in [³H]inositol incorporation into proteins. Wild-type (WT) and gpi11:: LEU2-pGAL-GPI11 (Δgpi11 + pGAL-GPI11) cells were maintained on SGlyYE medium, shifted to SGalYE or SGlcYE medium for 16 h, then resuspended in 1 ml of inositol-free medium containing Gal or Glc, respectively, and radiolabeled with 100 μCi of [³H]inositol for 60 min at 30°C. Proteins were then extracted and separated by SDS-PAGE, and radiolabeled GPI-anchored proteins were visualized by fluorography.