Figure 9. In vivo RNAi against TbAS-A in mice undergoing E. coli L-asparaginase treatment.

Time course of the parasitemia of two groups of 4 mice infected intraperitoneally, with 1×10⁴ of RNAi induced [using 1 mg/ml of doxycycline, Dox (+)] wt parental cell line (A) or a representative AS RNAi cell line (D) either untreated (open squares) or treated daily every 2 days (open diamonds) with 50 IU E. coli L-asparaginase/mouse. Parasitemia was quantified in peripheral tail blood at the times indicated. Error bars indicate standard deviation, and data are representative of two independent experiments using two different AS RNAi clones. The detection limit for this assay is 5×10⁴ trypanosomes/ml of blood. Mice were culled when parasitemia reached 1×10⁸ cells/ml. Kaplan–Meier survival plot of mice infected with RNAi induced wt cell line (B) or AS RNAi cell line (E) either untreated (open squares) or treated (open diamonds) with E. coli L-asparaginase. (F) Western blot analysis of AS-A levels in trypanosomes isolated from mice, before being euthanized. (C) Wt cell extracts, also collected when mice were culled, were use as negative control.