Transient TGFβ exposure causes persistent transdifferentiation in CDβGeo cells. (A) Description of experimental timeline – Cells were cultured and passaged in control media for 28 days (CDβGeo control) or 5 ng/ml TGFβ for 14 days followed by passage in control media for an additional 14 days. (B) The parental CDβGeo cells retain a cuboidal cobblestone morphology throughout the treatment period in the control media, but during TGFβ-treatment cells become senescent, spindle-like and avoid adhesion. (C) Western blot analysis shows down-regulation of E-cadherin and up-regulation of N-cadherin (D) Quantification of E-cadherin and N-cadherin protein expression, normalized to β-actin (* p < 0.05; **p < 0.01). (E) Immunofluorescence for E-cadherin and Actin demonstrate appropriate expression in control cells. TGFβ-treated cells have reduced E-cadherin expression and actin stress fibers. (F) K8 expression is reduced in TGFβ-treated cells.