Extended Data Fig. 7: The activation of RTK feedback that is induced after treatment with AZD8055 is disrupted in XL413-induced senescent cells.

a, Control cells and XL413-treated Hep3B cells were treated with AZD8055 for 48 h, and extracted proteins were analysed using a human phosphorylated-RTK array kit (left). The levels of phosphorylated RTK proteins were normalized to positive controls (right). b, The activation of RTKs identified by RTK arrays and the phosphorylation of SHP2 were validated by western blot analyses. c, Hep3B cells were treated with AZD8055 for 48 h before extraction of mRNA, and quantification of the indicated genes for RTK proteins was performed by qRT–PCR. Graph represents mean ± s.d. from three technical replicates. d, Hep3B cells were treated with AZD8055, and cell lysates were collected at the indicated time points to perform western blot analyses with the indicated antibodies. e, Hep3B cells were exposed to increasing concentrations of the AKT inhibitor MK-2206 in combination with AZD8055, and long-term colony-formation assays were performed; this revealed the synergistic effects of these two compounds on cell viability. f, Hep3B cells were treated with AZD8055, MK-2206 or a combination of both compounds at the indicated concentrations for five days, and apoptotic cells were visualized by caspase-3 and caspase-7 apoptosis assay. For gel source images, see Supplementary Fig. 1. All experiments shown (except for the RTK array analyses) are representative of two independent biological experiments.