Fig. 5. Phosphatidic acid generation by PLD influences the distribution of Asp5Ala(1-25)-4A1-GFP. (A) Cos1 cells were transfected with either (1-25)-4A1-GFP or the mutated construct D5A(1-25)-4A1-GFP. Cells were then treated with 0.3% (40 mM) butan-1-ol or 0.3% (40 mM) butan-2-ol or calphostin-c (1 µM) for 10 minutes at 37°C. (B) Cos1 cells were transfected with a construct of full-length PDE4A1, which was mutated at Asp5 to Ala. Cells were then treated with 0.3% (40 mM) butan-1-ol or 0.3% (40 mM) butan-2-ol for 10 minutes at 37°C, fixed in 4% paraformaldehyde, permeabilised and then stained for PDE4A. Cells were visualised using Alexa Fluor 594.