Figure 7.

Increased targeting of Gas1* to ERAD in GPI anchor lipid remodeling mutants. (A) CHX shut-off experiments with wild-type cells and remodeling mutants expressing HA-Gas1*. The lower part of the gel was separately stained with Coomassie as loading control. (B) Live cell fluorescence microscopy with wild-type cells and remodeling mutants expressing GFP-Gas1*. DIC = Nomarski image. Bar, 3 µm. (C–F) CHX shut-off experiments with the indicated double mutants expressing HA-Gas1*. The lower parts of the gels were separately stained with Coomassie as loading control. For quantification and statistical analysis, results from experiments shown in A as well as from those shown in Fig. 3 E (Δhrd1 and Δemp24 cells) were used. Mean values and SDs from at least three individual experiments are shown. Red circles are used to highlight the degradation rates in the double mutants.