Fig. 3. Nuclease activity of Leishmania endoG. (A) Western blot showing the reactivity of antibodies against HA with lysates of promastigotes (Pro) and axenic amastigotes (AxAm) of transfectants overexpressing the Leishmania endoG-HA protein. (B) Immunoprecipitated material (equal volumes) with antibodies against HA from KS, EG and EGm axenic amastigote cell lysates were resolved by SDS-PAGE and the blot was probed with biotinylated antibodies against rTbEG. (C) The immunoprecipitated material shown in B was incubated with 1 µg of plasmid DNA, and the cleavage products were resolved on a Bioanalyzer. The numbers on the x-axis represent the size of the DNA fragments in base pairs (bp). The peaks below 10380 bp in the LdEG and rTbEG panels represent cleavage products. rTbEG was used as a positive control, and KS controls (parasites transfected with the pKSNEO plasmid alone) are shown in A, B and C.