| Products/Services Used | Details | Operation |
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| Recombinant Proteins | Parasites were fixed with 3.8% (W/V) formaldehyde in PBS at 4°C, permeabilized with fresh PBS-0.1%Triton X-100 and blocked for 1 h at room temperature. PAR was detected with 1:500 primary anti-PAR antibody (BD) followed by 1:600 Alexa Fluor 594 goat anti-rabbit IgG antibody (Invitrogen). TbPARP was detected with 1:100 primary anti-TbPARP antibody (GenScript) followed by 1:600 Alexa Fluor 488 goat anti-rabbit IgG antibody (Invitrogen). Excess of antibody was removed by 3×5-min washes in PBS, and nuclear and kinetoplast DNA were stained with DAPI (2 μg/mL) (Sigma) in PBS. Coverslips were washed with distilled water and mounted in Mowiol and then visualized using an Olympus BX41 microscope. | Get A Quote |
Previously we demonstrated that an excess of poly (ADP-ribose) in the nucleus makes procyclic parasites more sensitive to hydrogen peroxide. However, the effect of an altered-PAR metabolism under standard conditions has not been addressed yet. Here we have analyzed the behavior of the growth curve of transgenic parasites that present this phenotype and studied cell cycle progression in synchronized cultures by flow cytometry and immunofluorescence. We have demonstrated that an excess of nuclear poly (ADP-ribose) produces a delay in the G1 phase of the cell cycle. Moreover, for the first time we have shown that poly (ADP-ribose) occurs at specific points very close to the mature basal body, suggesting there coul... More
