Fig. 2

Phenotypic characterization of HBEC CDC6 Tet-ON system during ECEE. a Ultrastructural analysis of 6 days induced cells revealed flattened cellular morphology, with enlarged nucleus (N), enhanced protein-synthesis, as visualized by double nucleoli (n) and extensive Golgi apparatus (G), and formation of extracellular vesicles (v). Arrows depict extracellular vesicles. Scale bars 2 μm and 500 nm. b IF analysis of the proliferative marker (Cyclin A) and CDC6 showed double positive cells only after escape from senescence. Scale bar: 15 μm. c IF for an epithelial marker (E-cadherin) or a mesenchymal marker (Vimentin) along with CDC6 indicated EMT in the “escaped” cells. IF for CDC6 showed ubiquitous and uniform expression in the induced cells (“ON” and “ESCAPED”). Scale bar: 20 μm