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Rosiglitazone-treated human umbilical vein endothelial cells were analyzed for growth rate by use of cell number counting, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay as well as 3H-thymidine incorporation.
The calibration curve between the DNA and cell number was prepared by use of cell suspensions with different cell densities.
Quantification of nucleosome concentrations in serum was done by use of Cell Death Detection Elisaplus of Roche Diagnostics (Mannheim, Germany).
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This has been achieved by use of cell-surface markers and by tagging cells which exhibit characteristics associated with stemness.
Cell-specific expression has been achieved by use of cell-specific promoters, intersectional promoters, and inducible strategies (Bacaj and Shaham 2007; Wei et al. 2012), some of which also enable temporal control of transgene activity.
Here, we have established a 3D cell culture model resembling the colonic crypt by use of HKe3 cells, human CRC HCT116 cells disrupted at activated KRAS.
By use of a cell line expressing the actin bundling WLIM domain under control of an inducible promotor we can activate membrane stabilization by the glucocorticoid analog dexamethasone.
The protocol includes a detailed description of a method for determination of the specific activity of the generated proteins by use of a cell culture based luciferase approach.
So far, the pathogenicity of AIEC has been studied mainly by use of cultured cell lines.
These colonies were expanded over several passages by use of standard cell culture procedures.
BrdU elisa assay was performed by use of a Cell proliferation Brdu Elisa Assay kit (Roche, 11647229001).
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