Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 3 - Phylogenetic tree of the DXS protein. The full set of DXS protein sequences annotated were obtained by BLASTP (Altschul et al., 1997) searched against the most recent versions of the non-redundant database at NCBI [http://www.ncbi.nlm.nih.gov/BLAST]. Protein alignments were made using ClustalW (Thompson et al., 1994) with no adjustment of the default parameters. Distance tree was constructed to maximum-parsimony with the PAUPSEARCH algorithm on Wisconsin Package Version 10.2, Genetics Computer Group (GCG), Madison, Wisc. [http://www.accelrys.com/about/gcg]. Numbers close branches indicate percent support in the bootstrap analysis (1,000 replicates).
• Supplemental Figure 1 - . Linearity of the antibody reactions. Linearity in protein analysis was corroborated by western blots testing serial dilutions of the same total protein extracts used for the 18-d-old ispD albino mutant (A) and 6-d-old Ler wild-type (B). Total protein concentration corresponds to 20, 10, 5 and 2.5 μg as indicated. Immunoblots were performed with antibodies raised against all MEP proteins, using the same dilution as specified in Methods.
• Supplemental Figure 2 - Comparative graphs of the densitometric quantification for the MEP proteins in response to the fosmidomycin treatment. The DXS protein level from the samples treated without (Fos−, dashed line) or with (Fos+, solid line) was obtained by densitometric analysis at each time point. The densitometric quantification was normalized to the level of an arbitrary protein from the coomasie blue-stained gel (arbitrary units) and referred to the DXS protein level at time 0 which was taken as 1. The data on the graph correspond to the mean ± SE of three independent experiments.