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In the culture at the low seeding density, an increase in confluence degree accompanied a decrease in growth potential (Rp), being Rp = 0.85 and 0.11 at t = 48 and 672 h, respectively.
An increase in confluence due to cell spreading and/or cell number was regarded as a trophic influence.
Several independent experiments showed a significant increase in confluence after addition of ACh (10 μM; Figures 1a and b).
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CD26 mRNA and protein levels increase in a confluence-dependent manner in colon carcinoma cell lines, with c-Myc acting as a repressor and Cdx2 acting as an enhancer of CD26 expression.
The marked increase in globosides at confluence depresses the relative abundance of other decorated subtypes, as shown in Fig. 6, after which there is a steady increase in GSLs decorated with fucose or sialic acid.
The cell proliferation is expressed as an increase in percentage of confluence.
Our data from Western blot analysis show that USF-1 protein expression is increased in a confluence-dependent manner, while HNF-1α expression is unchanged for both cell lines, suggesting that only USF-1 is correlated with the confluence-dependent increase for CD26 expression.
A potential explanation for the observed confluence-dependent increase in Cdx2 protein expression may be due to confluence-mediated cell cycle arrest, resulting in decreased expression of Cdk2 and decreased phosphorylation of Cdx2 by Cdk2, leading to increased Cdx2 protein expression [ 25].
The proliferation rates of the cell lines were determined by live-cell imaging, and are presented as increased cell confluence (in %) over time in Figure 4. Most of the cell lines proliferated very fast, reaching 60% confluence within 72 h when seeded at 5% confluence.
Only during the fifth day of culture i.e. when control and DEM+GSHe treated cells reached confluence, an increase in the number of dead cells occurred, but not in the DEM and BSO treated cells.
With increase in the overall clustering proportion confluence of individual clusters to larger modules is observed and, finally, large-scale aggregates, such as the tonotopic organization, are seen for maps with high clustering proportion.
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