Figure 2

Induction and cellular localization of Fmp45p-GFP. (A) Spectral cube (fluorescence) images were acquired using the Nuance™ Multispectral Imaging System (CRi) to assess cellular localization of Fmp45p-GFP and Sur7p-YFP in the multi-labelled strain SUR7-YFP FMP45-GFP. Individual localization is shown for each protein of interest (Sur7p-YFP and Fmp45p-GFP). Sur7p-YFP was artificially rendered in red so that co-localized proteins can be readily distinguished (yellow) in the merged image. (B) Localization of Fmp45p-GFP in either the wild-type (BWP17) or sur7 Δ null (SMB3) background was visualized by laser scanning confocal microscopy. Strains were grown at 42°C at a starting OD₆₀₀ of 0.1 in complete synthetic medium, supplemented with 1.0 M NaCl where required. After 24 h growth, confocal fluorescence images were documented using parameters optimized for imaging the sur7 Δ FMP45-GFP strain (sΔ-FMP45gfp) grown in the presence of high salt. Panels show fluorescence and DIC images of strains B-FMP45gfp (I and III, excluding and including salt, respectively) and sΔ-FMP45gfp (II and IV, excluding and including salt, respectively).