 Research articleA systematic approach to understand the mechanism of action of the bisthiazolium compound T4 on the human malaria parasite, Plasmodium falciparumKarine G Le Roch1 1 Department of Cell Biology and Neuroscience, University of California, Riverside, 900 University Avenue, Riverside, CA, 92521 USA 2 Department of Cell Biology ICND202, the Scripps Research Institute, 10550 North Torrey Pines Rd, La Jolla, CA, 92037, USA 3 Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR 5235, Université Montpellier II, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France 4 Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA, 92121, USA 5 Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030-3301, USA
BMC Genomics 2008, 9:513doi:10.1186/1471-2164-9-513
Additional filesAdditional file 1: Additional Figure 1: RT-PCR validation – RT-PCR experiments were performed to validate the microarray results. Plasmodium RNA was extracted at 24, 30, and 36 hours after 40 nM T4 treatment of synchronized cultures. Untreated RNA was extracted in tandem as a control. cDNA was created from the RNA extractions and quantified using spectrophotometry. 15 ng of cDNA were loaded into each RT-PCR reaction and was subjected to 20–35 cycles of amplification. Semi-quantifications of RT-PCR were done using NIH's ImageJ and illustrated by a column graph. Another column graph depicting the microarray results for the respective genes is shown for comparison. Pf14_0708, Mal13P1.129 and Mal7P1.100 were selected as genes expressed in gametocytes and showing a significant induction when incubated with T4); PF11_0509 and Mal7P1.6 showed an arrest of the cell cycle progression. Mal13P1.86, a gene involved in the parasite lipidic pathway is expressed and does not show any particular transcriptional change when incubated with the drug. Format: PDF Size: 99KB Download file This file can be viewed with: Adobe Acrobat Reader Additional file 2: Additional table 1: Transcriptional intensities at 30 minutes and 5 hours time points for treated and untreated parasite at 125 nM of T4 on a mix parasite population. Format: XLS Size: 863KB Download file This file can be viewed with: Microsoft Excel Viewer Additional file 3: Additional table 2: Transcriptional intensities at 24, 30 and 36 hours time points for treated and untreated parasite at 40 nM on synchronized parasites. Format: XLS Size: 1.1MB Download file This file can be viewed with: Microsoft Excel Viewer Additional file 4: Additional table 3: Known and putative phospholipid and neutral lipid enzymes of P. falciparum. Format: XLS Size: 35KB Download file This file can be viewed with: Microsoft Excel Viewer Additional file 5: Additional table 4: Protein abundance detected at 24 hours for treated and untreated parasite at 40 nM of T4 on synchronized parasites. Format: XLS Size: 209KB Download file This file can be viewed with: Microsoft Excel Viewer Additional file 6: Additional table 5: Nine proteins selected for the Targeted MS/MS approach. Format: XLS Size: 17KB Download file This file can be viewed with: Microsoft Excel Viewer |
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