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Bottom: fraction of solutions which are stable.
The most abundant functional class of genes in the CV TB bottom fraction encoded extracellular matrix (ECM) proteins (n = 18), indicating that basal taste bud cells may actively secrete matrix components and participate in basement membrane formation.
A total of 159 TB-associated genes exhibited intra-TB expression differences (≥5-fold CV TB bottom versus CV TB top ratio or ≥5-fold CV TB top versus CV TB bottom ratio, p value ≤0.05): 79 genes in the CV TB bottom fraction (Table S3) and 80 genes in CV TB top fraction (Table S4).
After centrifugation for 18 h at 100,000 g, gradients were fractionated from the top into five fractions of equal volume (with the solubilized pellet as the bottom fraction) and analyzed by SDS-PAGE, using a Tris-Tricine buffer system [65] and phosphorimaging using a Storm 840 phosphorimager and allied software (GE Healthcare Life Sciences, Piscataway, NJ).
In addition, ADAM10 was detected in the bottom fraction of the gradient.
In a subset of samples, polymorphonuclear neutrophils (PMN) were collected from beneath the Ficoll-Hypaque™ Plus layer (bottom fraction).
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In CC erythrocyte samples, we also detected most band 3 proteins in the bottom fractions 30 36.
These reductions in band 3 signal were observed in both in DRM and bottom fractions.
As shown previously for AA erythrocytes [34], [35], spectrin and glycophorin were found exclusively in the bottom fractions of CC erythrocyte preparations (Fig. 3).
Very-low and intermediate-density lipoproteins were removed, and the density of the bottom fractions was adjusted to 1.050 g/ml with solid KBr.
In contrast, signal distributions for both band 3 and CD47 in the bottom fractions were essentially indistinguishable (Figs. 4B and 4D).
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