Fig. 3.

Subcellular localization of LiTAT. (A) Confocal images were obtained using L. infantum early-logarithmic promastigotes and the green fluorescence corresponds to the Alexa Fluor 488 coupled to the primary antibodies specifically raised against the recombinant TAT. The blue fluorescence corresponds to the nuclear (N) and kinetoplast (K) DNA, stained with DAPI. LiTAT is distributed non-homogenously throughout the entire cytoplasm in early-logarithmic phase promastigotes. In stationary phase promastigotes any green fluorescence could be detected. (B) 10 μg of total protein extracts of selected fractions resulting from the digitonin gradient were subjected to Western blot analysis and probed with specific polyclonal antibody anti-LiTAT and anti-L. mexicana gGAPDH. Digitonin gradient assay allowed suggesting that the protein is cytoplasmic since the compartmentalized control protein (gGAPDH) is released to the supernatant with higher concentration of detergent than LiTAT as depicted in the graphics. (For interpretation of the references to colours in this figure legend, the reader is referred to the web version of this paper.)