Extended Data Fig. 10: SASP-mediated bystander effect with BRD inhibitor and DR5 agonist combination is regulated by NF-kB signaling.
From: cFLIP suppression and DR5 activation sensitize senescent cancer cells to senolysis
a, The proliferating A549 cells were cultured with SASP medium from different treatment-induced senescent A549 cells and treated with combination of 0.25 µM NEO2734 plus 4 µg/ml conatumumab or 1 µM navitoclax. b, A549 cells were cultured with different concentrations of SASP media and treated with the combination of 0.25 µM NEO2734 plus 4 µg/ml conatumumab. c, in vitro competition assay, to study the bystander effect of conatumumab plus NEO2734 in a heterogeneous population containing proliferating and senescent cells. GFP and RFP cells were generated through infection with a lentiviral vector coding for H2B-GFP and H2B-RFP. RFP labeled cells were induced to senescent by one week of 0.5 µM alisertib treatment. GFP labeled cells were used as proliferating cells. Different ratios of GFP and RFP cells were initially mixed and treated with the drug combination. The gray scale bars present 100 µm. d, RNA sequencing was performed on A549 cells cultured with SASP medium for 1 week and followed by gene set enrichment analysis (GSEA) of SASP medium cultured cells versus ctrl medium cultured cells for multiple independent NF-KB signaling gene sets (N = 1 RNA-sequencing data). e, P65 knockdown A549 cells were cultured with SASP medium and treated with 0.25 µM NEO2734 plus 4 µg/ml conatumumab. Error bars in this figure panel a-c, e represent as mean ± standard deviations, N = 3 independent experiments, statistical significance was calculated by two-tailed t-test.
