Fig 2.

(A) Functional activity of TP53 variants expressed in H1299 cells as determined by transactivation of TP53-responsive luciferase reporter (or a control reporter with point mutations in the response element which abolish TP53 binding [MutBS], Data Supplement). Expression of TP53 was confirmed by Western blot. C275X and Ins933a variants result in truncated proteins with faster migration. Vinculin staining demonstrates equal protein loading. All lanes except C275X and Ins933a demonstrate a full-length TP53 (53 kDa) and a smaller breakdown product. Loss-of-function alleles are associated with higher expression (presumably mediated by lack of negative feedback). (B) Data in panel A ranked based on activity and correlated with clinical presentation. Black rectangles indicate a family history consistent with Li-Fraumeni syndrome (LFS)/Li-Fraumeni–like syndrome (LFLS). White rectangles indicate absence of a significant family history in a complete pedigree containing three or more generations. Hatched rectangles indicate insufficient data to establish or exclude a family history of cancer predisposition. MPM, multiple primary malignancies; MR ACC, multiply recurrent adrenocortical carcinoma; mutBS, mutated TP53 binding site; WT, wild type.