Fig. 7. Effect of environmental temperature on the size and composition of the SMAFs in GFP8 cells. Cells, previously maintained at 15°C, were grown for 48 hours at the temperatures indicated. (A) GFP fluorescence images of cells grown at 15, 25 or 32°C. Bar, 5 µm. (B) Western blot analysis of control and GFP8 cells cultivated at 15, 25 or 32°C with anti-SFA. The amount of SFA-GFP decreased with rising temperatures, whereas slightly more SFA was detected in cells cultivated at 32°C. The amount of SFA in control cells remained almost constant under the conditions chosen. (C) Comparison of insoluble and soluble fractions from GFP8 cells cultivated at 25 or 32°C, revealing higher levels of SFA in the pellet at elevated temperature. Note the constant amount of SFA-GFP in the supernatant of GFP8 cells cultivated at 25 or 32°C. Anti-SFA was used to detect wild-type and GFP-tagged SFA. Similar amounts of whole cells, pellets and supernatants were loaded.