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. 2012 Dec 21;7(12):e52612. doi: 10.1371/journal.pone.0052612

Figure 6. Leishmania amastigotes lacking PIWI are more sensitive to apoptosis-like cell death inducing agents.

Figure 6

(A) LinPIWI(−/−) was shown to be highly sensitive to miltefosine (MF). Equal number of L. infantum axenic amastigotes (Ama) of wild type and LinPIWI(−/−) mutant strains were treated with various concentrations of MF for a period of 8 hrs. Drug sensitivity was evaluated by measuring the OD at 600 nm in a 96 well plates. The results are expressed as mean ± SD of biological triplicates and the experiments were conducted twice with similar results. The comparison of growth percentage between L. infantum WT treated with 0, 5, 10, 15, 20 µM of MF and LinPIWI(−/−) treated with 0, 2, 4, 6 and 8 µM of MF, respectively (LinPIWI(−/−) is much more sensitive to MF than WT) was analyzed by t-test (non parametric) using GraphPad Prism (version 3.03) software. Significant differences between WT and LinPIWI(−/−) are indicated (*, P<0.05; ***, P<0.001). (B) Primer extension analysis of L. infantum wild type (LinPIWI(+/+)) and five independent clones (Cl 1–3, Cl 5 and Cl 6) of LinPIWI(−/−) null mutant grown as axenic amastigotes using a forward primer corresponding to nucleotides 101–118 of the sLSU γ rRNA to detect antisense (as) LSU γ rRNA fragmentation.