Figure 9
The translation rate of the L. infantum HSP83 fluctuates during promastigote-to-amastigote differentiation. Wild-type (A) or pXIRPstCATIR-transfected (B) promastigotes were incubated during 0, 3, 9 and 24 h in amastigote growth conditions. The parasites were metabolically labeled with [35S]methionine/cysteine for the last 2 h of treatment. Proteins were extracted and analyzed by 10% SDS-PAGE, followed by autoradiography of the dried gels (panels a). The HSP83 band is marked with an arrowhead and the CAT band is marked with an arrow. The numbers at the left indicate the mobility and relative molecular weights (in kDa) of protein molecular weight standards. In parallel, cells were lysed and used to immunoprecipitate either the HSP83 (A) or CAT (B) using specific antibodies. Panels b show the immunoprecipitations followed by autoradiography; panels c show the immunoprecipitations followed by Western blotting. The label intensities of the immunoprecipitated proteins (panels b) were measured by densitometric scanning and normalized relative to the total amount of immunoprecipitated proteins (panels c). The relative signals are shown in panels d. Data represent mean values derived from at least three independent experiments.