At the late trophozoite stage synchronized parasites were treated with 0.5μM Shld1 or ethanol solvent control for 36h. Proteins were sequentially extracted as previously described2 (link), separated on 4-12% polyacrylamide gels (Life Technologies), and assayed using anti-HA tag (Roche Diagnostics 11 867 423 001), anti-Histone 3 (Abcam ab1791), and anti-PfPP2c (generous gift from C. Ben Mamoun). Replicates were biological not technical.
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Quantifying Protein Levels in Synchronized Parasites
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Corresponding Organization :
Other organizations : Pennsylvania State University, Princeton University, Fred Hutch Cancer Center, Universitat de Barcelona, University of London, Institute for Research in Biomedicine, Wellcome Sanger Institute, London School of Hygiene & Tropical Medicine, Institució Catalana de Recerca i Estudis Avançats
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Variable analysis
independent variables
- Treatment with 0.5μM Shld1
- Treatment with ethanol solvent control
- Protein expression levels (measured using anti-HA tag, anti-Histone 3, and anti-PfPP2c)
- Synchronized late trophozoite stage parasites
- Protein extraction and separation on 4-12% polyacrylamide gels
- Positive control: Not specified
- Negative control: Ethanol solvent
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