Extended Data Fig. 1: Models for oncogene- and therapy-induced senescence.
From: COPI vesicle formation and N-myristoylation are targetable vulnerabilities of senescent cells
a, IMR90 ER:RAS model for oncogene-induced senescence (OIS). Addition of 4-OHT activates ER:RAS, triggering OIS. b, Quantification of IF staining for BrdU incorporation in IMR90 ER:RAS cells 5 days after treatment with 4-OHT or DMSO (n = 3). c, Percentage of cells staining positive for SA-β-galactosidase activity in IMR90 ER:RAS cells 5 days after treatment with 4-OHT or DMSO (n = 3). Representative brightfield image (right). Scale bar, 100 µm. d, Percentage of cells staining positive for p16INK4a by IF in IMR90 ER:RAS cells 5 days after treatment with 4-OHT or DMSO (n = 3). Representative IF image (right). Scale bar, 100 µm. e, B-score normalization of cell numbers shown for the siRNA screen performed on control, non-senescent (DMSO treated) IMR90 ER: RAS cells. Points show normalized values for 3 technical replicates. Red box indicates samples with a B-score < 3. f, Screen results for control (DMSO-treated IMR90 ER:RAS) and OIS (4OHT-treated IMR90 ER:RAS). Graph displays B-score in OIS versus the negative of the difference in B score between OIS and control screens. Points show mean normalized values for 3 technical replicates with cut-offs shown for OIS B-score < -3 and a difference in B score of > 2 between control and OIS cells. g, Model of therapy-induced senescence (TIS). Senescence was induced by 7 days of doxorubicin treatment in IMR90 cells. h, Quantification of IF staining for BrdU incorporation in IMR90 cells 2 days after treatment with doxorubicin or DMSO (n = 3). i, Percentage of IMR90 cells staining positive for SA-β-galactosidase activity 7 days treating with doxorubicin or DMSO (n = 3). Representative brightfield images (right). Scale bar, 100 µm. j-k, Percentage of IMR90 cells staining positive for p21CIP1 (j) or p16INK4a (k), 6 (j) or 3 (k) days by IF after treating with doxorubicin or DMSO (n = 3). Representative IF image (right, j & k). Scale bar, 100 µm. Data represented as mean ± SD unless otherwise stated. Statistical tests performed using unpaired two-tailed t-test unless. N represents independent experiments. Source numerical data are available in source data.
