Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in -0

Non-irradiated (No UV-C) and irradiated (UV-C) trophozoites harvested at different times (30 min, 3, 6 and 12 h). Upper panels, histograms show the DNA fragmentation percentage in fluorescence positive cells. The abscissa indicates fluorescence of propidium iodide (PI), and the ordinate indicates fluorescence of Alexa 488-labeled 3' ends of DNA. The number inside each histogram denotes the percentage of fluorescence positive cells above the cut-off line. Lower panels, PI-staining cells were checked in the epifluorescence microscope to confirming the absence of cytoplasmic stain. PI, propidum iodide, N, Nomanski optics. . Neutral comet assays of non-irradiated (No UV-C) and irradiated (UV-C) trophozoites harvested at different times (30 min, 3, 6 and 12 h). Electrophoretic migration of DNA was from left (anode) to right (cathode).<p><b>Copyright information:</b></p><p>Taken from "Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in "</p><p>http://www.biomedcentral.com/1471-2199/9/35</p><p>BMC Molecular Biology 2008;9():35-35.</p><p>Published online 10 Apr 2008</p><p>PMCID:PMC2324109.</p><p></p

Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in -6

Ee probe. Lanes 2 to 4, ssDNA incubated with increasing amounts of rEhRAD51 (2.5, 5 and 7.5 μg, respectively); lanes 5 to 7, ssDNA incubated with increasing concentrations of mock purified fraction (2.5, 5 and 7.5 μg) as control. Protein-DNA complexes (Cto C) are shown by arrowheads. . Partially purified rEhRAD51 was incubated with [α-P]dATP labeled dsDNA and interactions were resolved through PAGE. Lane 1, free probe. Lanes 2 to 4, dsDNA incubated with increasing amounts of rEhRAD51 (2.5, 5 and 7.5 μg, respectively); lanes 5 to 7, dsDNA incubated with increasing concentrations of mock purified fraction elution fraction (2.5, 5 and 7.5 μg) as control. Protein-DNA complexes (Cto C) are shown by arrowheads. . D-loop reactions containing 10,000 cpm of [γ-P]dATP-labeled oligonucleotide, circular dsDNA and 0, 2.5, 5 and 7.5 μg of partially-purified rEhRAD51 (lanes 1 to 4) were incubated at 37°C for 30 min with 2 mM of ATP. Negative controls were performed without homologous dsDNA (lane 5) and with heterologous dsDNA oligonucleotide instead of homologous dsDNA (lane 6), both of them using 7.5 μg of EhRAD51 elution fraction. Reaction products were analyzed by agarose gel electrophoresis, transferred to nylon membranes and visualized through a Phosphor Imager. . Densitometric analysis of D-loop products obtained in C. Results are representative of two independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in "</p><p>http://www.biomedcentral.com/1471-2199/9/35</p><p>BMC Molecular Biology 2008;9():35-35.</p><p>Published online 10 Apr 2008</p><p>PMCID:PMC2324109.</p><p></p

Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in -2

D trophozoites harvested at different times (UV-C; lane 2, 0.5 h; lane 3, 3 h and lane 4,12 h). Arrowheads denote the length (bp) of each expected amplified internal fragment, as described in Table 2. . Densitometric analyses of RT-PCR products in A. Pixels corresponding to the rRNA product were taken as 100% in each lane. Data are the mean of three independent assays.<p><b>Copyright information:</b></p><p>Taken from "Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in "</p><p>http://www.biomedcentral.com/1471-2199/9/35</p><p>BMC Molecular Biology 2008;9():35-35.</p><p>Published online 10 Apr 2008</p><p>PMCID:PMC2324109.</p><p></p