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The quantification of the intensity of the bands from western blots was made using Image J. The values included in the graph represent the average from 3 experiments after the normalization; in each case, we used pools of material from 4 animals per genotype and the data represent mean ± SEM.
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Southern blots were made using 20 µg DNA and the Vaku-Blot system (Biometra).
"Virtual northern blots" were made using the Clontech SMART cDNA Synthesis Kit, according to the manufacturer's instructions using RNA from the stages indicated.
Western blot analysis was performed using a chemiluminescence system (Luminol) and detection was made using a CCD-camera (LAS 1000, Fuji).
Testing for 44 HPV types was made using a general primer GP5+/6+ -mediated PCR and reverse-line blot hybridization [ 12].
Results for the microarray experiments were verified using "virtual northern blots" which were made using the Clontech SMART cDNA Synthesis Kit, according to the manufacturer's instructions using RNA from the same stages used in the microarray experiment.
To quantitate the GFP bands from the Western blot, densitograms were made using ImageJ software (http://imagej.nih.gov/ij/, last accessed April 30 , 2015.
Probe DNA for Southern blotting was made by amplification of DT40 genomic DNA using the primers cWRKOPF1 [5′-GTTAATACCGTGGCTTGCTGAAGCATTTCTTGAC] and cWRKOPR2 [5′-GCTCAGCTGTAGGCTTTTGTTTTAATGAACACAAC], cloned into pCR 2.1-TOPO then excised as a SpeI, XhoI fragment.
Analysis of blots was performed using ImageJ (National Institutes for Health).
Similar observations were made in our previously published study using western blotting [ 16].
Densitometric analysis of western blots was carried out using Vision Works software (version 6.7.1, Upland, CA, USA).
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