Extended Data Fig. 3: Cell killing is specific to p53-01.
From: Mutant p53 protein accumulation is selectively targetable by proximity-inducing drugs
a, Structure of KG5-PEG4-BI2536. b, Normalized nanoluciferase signal one day after dose titration of PMV6 and BI-2536 in 293 T cells co-transfected with NLS-LgBiT-p53Y220C(DBD) and mEGFP-PLK1-SmBiT. N = 4 technical replicates/condition. c, Live cell imaging of 293 T cells co-transfected with Halo-p53Y220CΔTAD-mCherry and mEGFP-PLK1-SmBiT and treated with DMSO, PMV6, or BI-2536 for 16 h. Two replicates of this experiment were performed; a representative replicate is shown. Scale bar is shown (10 μm). d, Crystal violet staining of 293 T cells stably expressing Halo-p53Y220CΔTAD-mCherry versus parental 293 T cells treated with PMV6 or BI-2536, analyzed after 5 days. Experiment performed in triplicate; representative replicate shown. e, Structure of PMV6-PEG4-adavosertib. f, Competition of Halo-p53Y220CΔTAD-mCherry vs. parental 293 T cells with p53-01 or PMV6-PEG4-adavosertib for 10 days; analyzed for mCherry expression by flow cytometry. N = 3 measurements/condition. g, Proportion of late apoptotic cells (Annexin+, PI+) following one day treatment with p53-01, BI-2536, or PMV6 at indicated concentrations in Halo-p53Y220CΔTAD-mCherry vs. parental 293 T cells. n.s. not significant, **P < 0.01, ***P < 0.001 by t-test. N = 3 measurements/condition. h, Caspase-3/7 glo of Halo-p53Y220CΔTAD-mCherry vs. parental 293 T cells treated with BI-2536 or PMV6 after one day. N = 4 technical replicates/condition. i, Normalized viability of 293 T cells expressing a dox inducible Halo-p53Y220CΔTAD-mCherry treated with p53-01, PMV6, or BI-2536 for 4 days, induced with dox at various concentrations the day before beginning compound treatment (0 ng/mL, 100 ng/mL, 1000 ng/mL). N = 4 technical replicates/condition.
