Fig. 1.

 Pdr10 localizes to punctate structures in the plasma membrane and to the detergent-resistant fraction. a Strain NRY912 was grown in YPD for 2 days, and Pdr10-GFP was examined by fluorescence microscopy. Pdr10-GFP function was confirmed by Calcofluor sensitivity. b Strain NRY912 was grown for 2 days as in (a) but was treated with latrunculin A prior to microscopy. Bar for (a) and (b), 6 μm. c Cells were repeatedly exposed to look for movement of Pdr10-GFP puncta (arrowheads). Shorter exposure times and higher gain were used to permit the maximum number of exposures prior to loss of signal due to photobleaching. Bar, 3 μm. d Total membranes were isolated from log phase cells of strain NRY930, extracted with cold Triton X-100, and analyzed by flotation though Optiprep density gradients with subsequent immunoblotting as described under Materials and Methods. Strain NRY930 expresses a PDR10 allele with a C-terminal tandem myc-His₆ (mh) tag. The function of the resulting Pdr10-mh protein was confirmed by Calcofluor sensitivity. Hxt1, Gas1, and Pma1 were used as markers for detergent-soluble and detergent-resistant membranes