Extended Data Fig. 9: Death receptor activation mediated immune surveillance and the evidence of | SASP-mediated bystander effect.
From: cFLIP suppression and DR5 activation sensitize senescent cancer cells to senolysis
a, Fixed tissues were dehydrated and embedded in paraffin. Sections of 2-4 µm were prepared and immunostained with P21 to indicate senescent cells in the tumors of A549. The black scale bars present 50 µm. b, the quantification of the P21 immunostaining (N = 18 tumor tissue slides). c, Transcriptomic analysis of CD95 (FAS) in A549 cells treated with 1 week of 0.5 µM alisertib. d, Dose-response curves in 0.5 µM alisertib-induced senescent A549 and SK-Hep1 cells treated with CD95 ligand (FASLG). e, Senescence associated beta-galactosidase staining on one week of different SASP media cultured A549 cells (the media were collected from the senescent models induced by different treatments). 0.5 µM Alisertib-induced senescent cells were used as the control. The black scale bars present 100 µm. f-g, SASP medium was collected from the alisertib-induced senescent cells and mixed with an equal volume fresh medium. Ctrl medium was collected from proliferating cells and mixed with an equal volume fresh medium. Afterwards, The SASP medium or Ctrl medium were added to new proliferating A549 (f) and SK-Hep (g) cells and treated with NEO2734 and Conatumumab. h, Cell viability accessed by colony formation of SASP medium cultured A549 cells treated with the combination of 0.25 µM NEO2734 plus 4 µg/ml conatumumab and 10 µM Z-VAD-FMK. The results in panel d, e represent at least 3 times with similar results. Error bars in this figure panel b represent for N = 18 tumor tissue slides. Results in panel f-h represent by N = 3 independent experiments. These results represent as mean ± standard deviations, statistical significance was calculated by two-tailed t-test.
