Figure 3.

Analysis of EpCAM target genes in HMECs. HMECs were cultivated as mitotic subconfluent cultures (A) or for 10 days on Matrigel-coated transwells to induce polarization of epithelial monolayers (B). Then, cells were adenovirally transfected (MOI = 100) to overexpress GFP alone or EpCAM/GFP. Polarized HMECs cells did not express immunoreactive EpCAM protein, but the gap junction protein ZO-1, E-cadherin and β-catenin as determined by Immunofluorescence analysis (C, bars indicate 25 μm). Polarized cells and mitotic cultures (log-phase) of three donors (HMEC 1–3) were adenovirally transfected and EpCAM gene expression quantified 24 h later by real time PCR using GAPDH as internal house-keeping gene (D). MA-plot of genes regulated by EpCAM as determined by Affymetrix Gene analysis in mitotic standard cultures (E) or confluent polarized cultures of HMECs (F). Besides EpCAM no additional genes were significantly regulated in all three donors analyzed 20 h after transfection. Mean M-values indicate differential gene expression between EpCAM over-expressing and GFP transfected cells (log2 scale), Mean A-values indicate average expression of a gene in all microarrays. Stars indicate p values <0.05.