Fig. 5
NMT is essential in L. donovani amastigotes: qualitative analysis. Representative flow cytometry plots (plotted y-axis: tdtom, x-axis: GFP) from parasites obtained from spleens of BALB/c mice treated for 28 days with 3 mg/kg per day− b.i.d with ganciclovir (+; GCV) or with solvent (−). The predominant population was identified for each condition (single or double knockout, transfected with NMT-TK plus either the NMT-tdtom or the tdtom plasmid) by selecting for no plasmid (medium only, wt; hygromycin NMTHYG/+ [NMT-TK]; all others hygromycin and puromycin) in the top two rows; for NMT-TK plasmid (as before plus nourseothricin; rows 3 and 4); for NMT-tdtom or tdtom plasmid (hygromycin, puromycin and G418; rows 5 and 6); for NMT-TK/NMT-tdtom or NMT-TK/tdtom plasmid (hygromycin, puromycin, nourseothricin and G418; rows 7 and 8). See Additional files 4, 5, 6, 7, 8: Figures S4–S8 for flow cytometry plots for each mouse. Flow cytometric analysis was performed 14 days after spleens were removed and parasites grew out from cell suspension