Notch1 activation and TGF-beta stimulation in transformed human esophageal cells. Homo sapiens

Transformed human esophageal keratinocyte cell line EPC2-T (EPC2-hTERT-EGFR-cyclin D1-p53R175H) cells were stimulated with or without 2.5 ng/ml recombinant human TGF-beta1 for 10 days. The above cells were subjected to treatment for 10 days with or without 0.5 µg/ml doxycycline (DOX) to activate tetracycline-inducible (tet-on) ICN1, an active form of Notch1. Overall design: Transformed human esophageal keratinocytes EPC2-T (EPC2-hTERT-EGFR-cyclin D1-p53R175H) were stably transduced with tetracycline-inducible ICN1 (active form of Notch1). Cells were treated for 10 days either with or without 2.5 ng/ml recombinant human TGF-beta1 (TGF) along with or without 0.5 µg/ml doxycycline (DOX). Then, cells were subjected to RNA extraction in triplicate for gene array experiments. Four conditions were compared: (1) neither TGF-beta1 treatment nor Notch1 activation (cont); (2) Notch1 activation only (DOX); (3) TGF-beta1 stimulation only (TGF); (4) Notch1 activation plus TGF-beta1 stimulation (DOXTGF).