Elevation of cellular PA redistributes Chm7-GFP to envelope/ER membranes (supplemental to Fig. 4 ). (A) Elevation of cellular PA levels leads to Chm7 recruitment to NE/ER membranes dependent on the hydrophobic stretch. Localization of Chm7-GFP or chm7-W3A1VA-GFP in the indicated strains with the NE and ER membranes visualized with dsRED-HDEL. GFP (inverted fluorescence), dsRED, and merged images are shown. Arrowheads point to colocalization of Chm7-GFP with NE/ER membranes. Scale bar, 2 µm. (B)CHM7 is required for fitness of pah1Δ but not dgk1Δ strains. 10-fold serial dilutions of the indicated strains grown on YPD plates at 30°C or 37°C for 48 h before imaging. (C)CHM7 is dispensable for the fitness of nem1Δ strains. 10-fold serial dilutions of the indicated strains grown on yeast extract peptone dextrose plates at the indicated temperatures for 48 h before imaging. (D) The NLS-DAG mCherry sensor does not focally accumulate at the NE in wild-type strains. Deconvolved inverted fluorescence micrographs of NLS-DAG sensor-mCherry at 23°C or at 37°C. Red asterisks demark nuclei. Numbers are mean percentage and SD of cells from three replicates of >50 cells per replicate with the appearance of NLS-DAG sensor-mCherry foci at the NE. Scale bar, 5 µm.