Extended Data Fig. 8: Treatment with AZD8055 does not induce apoptosis in cisplatin- or alisertib-induced senescent cells.

a, BJ/ET/RAS^V12 cells were treated with 100 nM 4-OHT for 21 days to induce senescence, as detected by SA-β-gal staining. b, Control or senescent BJ/ET/RAS^V12 cells were treated either with vehicle or with 400 nM AZD8055 for 96 h, and apoptotic cells were visualized by caspase-3 and caspase-7 apoptosis assay. c, Hep3B cells were cultured in the presence of cisplatin or alisertib (aurora-A kinase inhibitor) for 4 days at the indicated concentrations, and the induction of senescence was detected by SA-β-gal staining. d, Hep3B cells were treated with cisplatin (1 μg ml⁻¹) or alisertib (250 nM) for 4 days, and subsequently exposed to vehicle or 400 nM AZD8055 for 96 h. Apoptotic cells were visualized by caspase-3 and caspase-7 apoptosis assay. e, Control cells, or cisplatin-, alisertib- or XL413-induced senescent cells were treated with AZD8055, and cell lysates were collected at the indicated time points. Western blot analyses were performed with the indicated antibodies, which revealed that the mTOR signalling feedback loop is functional in cisplatin- and alisertib-induced senescent cells (whereas it is efficiently inhibited in XL413-induced senescent cells). For gel source images, see Supplementary Fig. 1. Data in a–e are representative of two independent biological experiments.