Figure 2
Expression of CAT along the cell cycle in the different transfected Leishmania cell lines. Protein extracts from 35S-labelled L. infantum promastigotes transfected with the indicated constructs and treated with 5 mM HU either for 12 h (lane 0) or at the indicated time period (in h) after removal of the drug, were used to immunoprecipitate CAT. After immunoprecipitation, protein samples were separated by SDS/PAGE and transferred on to a nitrocellulose membrane. After autoradiographic exposure (De novo panels), membranes were incubated with the anti-CAT antibody to reveal the total amount of CAT present in the samples (TP panels). Graphics show the ratio between de novo synthesis of CAT and total amount of CAT determined by densitometric quantification (columns) and the percentage of cells in the S phase of the cell cycle determined by flow cytometric analysis (line). Data correspond to one representative experiment of three independent assays with similar results.