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The biological significance of the double staining for each of the mentioned pairs (ET, cell marker) was determined by calculating the Pearson's correlation coefficient (rp) for each analyzed region of interest (ROI) in multiple slices.
The concentration of each marker was determined using a standard curve that was constructed by plotting the mean optical density obtained for each reference standard provided by the kit against the known concentration.
For Chromosome 8, the regression tree for marker D8S516 (Figure 4c) at the original "Hindu" peak of linkage showed that the primary binary decision for families contributing to linkage at this marker was determined by whether they lived east or west of the UTM easting 323,564 m.
For Chromosome 6, the regression tree for marker D6S244 (Figure 4a) at the original "Hindu" peak of linkage showed that the primary binary decision for families contributing to linkage at this marker was determined by whether they lived north or south of the UTM northing 2,886,078 m.
Similar results were also observed when serum pentosidine as another marker was determined.
The number of cells expressing a specific marker was determined as described in the ESM Methods.
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Genomic position and expected PCR product size for each marker were determined based on the reference sequence of rice cultivar Nipponbare released by International Rice Genome Sequencing Program IRGSPP).
In addition, CXCL-5, a chemotactic chemokine, and ICAM-1, an endothelial activation marker, were determined according to the recommendation of the manufacturer (R&D Systems, Wiesbaden, Germany).
The genotypes of IL-8 (-251) T/A marker were determined as previously reported [ 30].
The median, first and third quartile values for each molecular marker were determined using all informative samples.
The mRNA levels of PPAR γ, FABP4 and RUNX2 (used as a bone marker) were determined by qRT-PCR.
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