Fig 5. Extraction, quantification and size determination of extracellular polyP.

100mg of mycelium of the WT and phoP mutant, grown on a Pi limited solid R2YE medium, was washed with sterile water or with a solution of 5 mM EDTA. PolyP was extracted from the washing solutions, quantified and run on a on urea-polyacrylamide gel for size determination at different time points throughout growth. (A) The plain grey and striped grey histograms represent polyP found in the wash solution of the WT strain with water or with 5 mM EDTA, respectively. (B) The plain white and striped white histograms represent polyP found in the wash solution of the phoP mutant with water or with 5 mM EDTA, respectively. The error bars represent the standard deviation of three independent experiments. (C) Poly P collected after EDTA washing of 100mg of mycelium of the WT and phoP mutant strains after 80 h of cultivation was analyzed on a urea-polyacrylamide gel. Size standard polyP, approximate length 20, 40 and 60 residues, are shown in lanes 1, 2 and 3. The polyP is approximately 10 residues long in both strains and is more abundant in the WT than in the phoP mutant strain.