FIGURE 5:

Modulation of fat1Δ-induced phospholipid saturation by the desaturase Ole1. (A) Ole1 protein levels after GAL1-promoter shutoff. Whole-cell extracts were prepared at the indicated time points and analyzed by SDS–PAGE followed by immunoblotting. Top, anti-HA antibody; bottom, anti-Pgk1 antibody (loading control). (B) Relative intensities of PC (32:1 and 32:2) species in the indicated strains. For Ole1 knockdown , cells were cultured in glucose-containing media for 8 h (see A). Culturing in galactose-containing media actually resulted in Ole1 overexpression, as can be seen in the increased unsaturation seen in the GAL1-Ole1 strain vs. the wild-type strain (compare first and fifth samples). Relative intensity is expressed as a percentage of the combined intensities of the two molecules. Similar results were obtained in three independent experiments. These experiments were performed using LESA MS.