Extended Data Fig. 1: CRISPR screen optimization, validation, and YAP-TEAD inhibition in other senescence models.
From: The YAP–TEAD complex promotes senescent cell survival by lowering endoplasmic reticulum stress
a, Cell viability assessment by direct cell counting of senescent WI-38 cells treated with puromycin (1 µg/ml, 48 h) 72 h after being transduced with the Brunello library at the indicated MOIs. The gray bars represent the expected viability if the transduction efficiency was complete while the teal bars represent the viability observed for each of the MOIs after puromycin treatment. b, Cell viability as assessed by direct cell counting of WI-38 cells transfected with the indicated siRNAs and rendered senescent after treatment with etoposide for 6 days (ETIS). c, Analysis of the levels of the indicated mRNAs in proliferating (P) or ETIS WI-38 cells transfected with the indicated siRNAs 24 h before either treatment with etoposide (50 µM) or no treatment, and culture for an additional 6 days. d, Representative western blot analysis (n = 3 independent experiments) of the levels of phosphorylated YAP (S127), YAP, phosphorylated MOB1 (T35), MOB1, and ACTB levels at the indicated conditions. e,f, Analysis of BrdU incorporation (e) and SA-β-Gal staining (f) in the indicated cell types, rendered senescent by etoposide (ETIS), ionizing radiation (IRIS), or replicative exhaustion (RS). Scale bar 100 µm. g, Caspase 3/7 activity measured in RS and IRIS WI-38 cells treated for 72 h with the indicated doses of Verteporfin (VPF). h, i, Cell viability as assessed by direct cell counting (h) and Caspase 3/7 activity measurement (i) for the indicated models of senescence along with proliferating controls, after either no treatment or treatment with VPF for 72 h at the indicated doses. Graphs in (b, c, e, g–i) represent the means and each individual value as a dot ±s.d. n = 3 independent replicates; significance (*P < 0.05, **P < 0.01, ***P < 0.001) was determined using two-tailed Student’s t-test. Unless indicated, statistical tests were performed relative to untreated or proliferating controls.
