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. Author manuscript; available in PMC: 2018 Mar 23.
Published in final edited form as: Cell. 2017 Mar 23;169(1):132–147.e16. doi: 10.1016/j.cell.2017.02.031

Fig. 1. FOXO4 is elevated in senescent normal fibroblasts and ensures their viability.

Fig. 1

A) Schematic representation of the mRNA expression changes (Fig. S1A) of the cell-intrinsic apoptosis pathway(Tait and Green, 2010) between senescent and control (proliferating) IMR90 fibroblasts. Inset: Immunofluorescence for PUMA, BIM and BCL2. B) Volcano plot comparing transcriptional regulators in senescent vs. control IMR90. (See Fig. S1B for expression and p-values). Dark blue: associated with apoptosis. Inset, left: RNA expression of the FOXO cluster. N.D. Not detectable. Right: Protein levels of the FOXO cluster. FOXO1 was ectopically expressed as positive control. C) QPCR for changes in FOXO1, 3 and 4 mRNA after senescence-induction by 10Gy IR. p21Cip1 (biphasic increase), p53 and ETS2 (biphasic decrease) are included as controls. D) Immunoblot for changes in FOXO3 and 4 protein levels after senescence-induction by 10Gy IR. E) The senescence-induced FOXO4 mRNA expression is successfully countered by two shRNAs. F) Cytochrome-C release assay (left) as measure for apoptosis in the conditions of E), quantified in a histogram (right). G) Induction of cleaved Caspase-3 after senescence induction in (mouse-specific) FOXO4-deprived wildtype or bax/bak−/− BMK cells. H) AqueousOne viability (left) and colony density (right; see also Fig. S1C) of control and senescent IMR90 cells transduced with the short hairpins used in E).