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Integrated optical density (IOD) of each band was calculated using Image Master VDS software (Amarsham Biosciences, UK).
To quantify western blots, TIF images were recorded for each blot using a Chemidoc system (Bio-Rad), converted to 8-bit grayscale using Adobe Photoshop, and the integrated intensity of each GFP and α-tubulin band was calculated using ImageJ.
The mean score of those not offered a place was 627.5 (76.4%) (SD = 67.6 [8.2%]).The NAC total assessment score used to determine the AGPT ranking band was calculated using a Z-score (Total Score-Mean Score/Standard Deviation) and a T-Score (Z-Score*10 + 100) for each assessment measure.
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Five formulations of PRI (based on R531 as a xanthophyll-sensitive spectral band) were calculated using different reference wavelengths, such as PRI570 (reference band RREF = R570), and the PRI modifications PRIm1 (RREF = R512), PRIm2 (RREF = R600), PRIm3 (RREF = R670), and PRIm4 (RREF = R570, R670), along with other structural indices such as NDVI, SR, OSAVI, MSAVI and MTVI2.
To analyze the experimental results, all energy levels in the conduction and in the valance band were calculated using transfer matrix technique, taking into account the effective mass and the parabolic band approximations.
The average intensity of bands was calculated using ImageJ.
Quantification of bands was calculated using ImageJ (NIH), a Java-based image-processing program.
The intensity of the bands was calculated using Image J (http://imagej.nih.gov/ij/) on images taken using a G Box imager (Syngene, Cambridge, UK).
The amplified products were subjected to agarose gel electrophoresis, and the intensity of resultant bands was calculated using a Lane and Spot Analyzer (Atto, Japan).
The antibodies were considered specific based on previous data and detection of few bands in the blots, and in all cases the molecular weight of the corresponding bands was calculated using Quantity One software (BioRad).
Statistical significance of the mean IOD values for immunoblot bands was calculated using the two-way analysis of variance (two-way ANOVA) and Bonferroni multiple-comparison correction, when the comparison was made between the treatments (control and E2, P4, and E2 + P4) and duration of 3D culture (3, 6, 9, and 14 days).
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