Fig. 2

Anx family members translocate to the surface of apoptotic cancer cells. A Schematic representation of experimental procedure. Apoptosis in cells was induced by treatment with Etoposide (Eto), Dimethylfumarate (DMF) or UV-C radiation followed by incubation for 48 h. Subsequently, the cell surface was washed with 20 mM EDTA/PBS and supernatants were analysed by immunoblot. Washes of the same number of untreated live cells served as controls (-). B EDTA washes of indicated cancer cell lines before and after apoptosis induction were probed for Annexin family members (AnxA1-A13) by specific antibodies. Detection of cytosolic Erk1/2 was used as a control for cell membrane integrity. General protein content of the washes was indicated by Ponceau staining. Full-length blots are supplied in Supplementary Information Figures S7, S8, S9. In all experiments, the induction of apoptosis was monitored and confirmed by flow cytometry using AnxA5/7AAD staining (see Supplementary Information Figure S3).