Infection of mouse macrophages and BALB/c mice footpads with WT <i>L</i>. <i>braziliensis</i> and RNAi-1060 cell lines.

<p>(A) Intraperitoneal macrophages from BALB/c mice were incubated for 24 hours at 34°C with stationary phase promastigotes from WT <i>L</i>. <i>braziliensis</i> cultures and the two cloned cell lines RNAi-Ama1060 cl1 and cl5 at a ratio of 10: 1 parasites/cell. After washing non-internalized promastigotes macrophages were incubated for 24, 48 or 72 hours before the cells were stained with DAPI and the numbers of intracellular amastigotes, visualized by fluorescence microscopy, were determined. (B) BALB/c mice were infected in the footpads with 10⁷ stationary phase promastigotes from WT <i>L</i>. <i>braziliensis</i>, <i>L</i>. <i>braziliensis</i> transfected with the pIR1PHLEO vector containing GFP, and two cloned cell lines expressing δ-amastin siRNA, RNAi-1060cl1 and cl5. Two, four and nine weeks after infection, parasitism was evaluated by the limiting dilution method.