Figure 7.

elo3Δ erg6^ts affects steady state raft association of the plasma membrane ATPase. Raft association of Gas1p and the plasma membrane ATPase, Pma1p, in the elo3Δ erg6Δ double mutant strain bearing a plasmid-borne copy of either ERG6 (elo3Δ ERG6, YRS1520) or erg6^ts (elo3Δ erg6^ts, YRS1519). Cells were cultivated at 24°C or shifted to nonpermissive conditions (37°C) for 2 h, before isolation of detergent-resistant membranes by either one (1st Gradient) or two rounds (2nd Gradient) of detergent extraction and floatation on density gradients (Bagnat et al., 2000). The position of mature Gas1p and that of Pma1p is indicated. For the mixing experiment, cells of the two genotypes were mixed at a 1:1 ratio after incubation at nonpermissive conditions for 2 h, and detergent solubility of Gas1p and Pma1p was assessed after two successive rounds of detergent extraction and floatation on density gradients. All blots were processed under comparable conditions.