Figure 4

Time and temperature dependent accumulation of hERG-TEV-GFP-His ₈ . A) Exponentially growing cells cultivated at room temperature in expression medium until OD ₄₅₀ = 1.0 were separated in two. One half was transferred to 15°C while the other half was inoculated at 30°C. After 15 minutes of thermo-equilibration, production of hERG-TEV-GFP-His ₈ was induced by addition of Galactose (T = 0). Fluorescence was determined in duplicates of crude membranes isolated from yeast cells induced for the indicated periods of time at either 15°C (blue line squares) or 30°C (red line circles). Fluorescence was translated into pmol hERG protein/mg total membrane protein using a GFP standard curve. Standard deviations of duplicates are shown as error bars. B) In-gel fluorescence of 80 μg crude membranes prepared from the cultures induced at 15°C used in figure A. C) In-gel fluorescence of 80 μg crude membranes prepared from the cultures induced at 30°C used in figure A. Lanes are marked with time of hours post induction.