Cell Cycle Synchronization of Intracellular Parasites
Corresponding Organization :
Other organizations : Lanzhou Veterinary Research Institute, Shanghai Clinical Research Center
Variable analysis
- Compounds (5 μM each)
- Cell cycle synchronization of intracellular tachyzoites
- Intracellular tachyzoite population
- Monolayers of HFF (human foreskin fibroblasts)
- Tachyzoite infection at MOI = 1 (multiplicity of infection)
- Synchronization with 80 μM pyrrolidine dithiocarbamate (PDTC) for 8 h
- Incubation time of 8 h after compound treatment
- Purification of intracellular tachyzoites using a 27-gauge needle and 3-μm filter
- Fixation of parasites with 70% ethanol at -20°C for 24 h
- Staining with propidium iodide (PI) at 37°C for 30 min
- Filtering with a 5-μm pore filter
- Detection on FACSCanto™II flow cytometer
- Analysis using FlowJo 7.6.1 software
- Collection of at least 50,000 events per sample
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