Fig 2: Characterizing allosteric inhibitors of the CCNE1:CDK2 complex.a, Structures of two inhibitors of the CCNE1:CDK2 complex – I-125A and I-198 – reported in a patent26. b, ADP-Glo data showing effects of the indicated compounds on the activity of a purified N112C-CCNE1:CDK2 complex as measured by phosphorylation of the C-terminal peptide of RB1 (left) or the histone H1 protein (right). The N112C-CCNE1:CDK2 complex was pre-treated with compounds for 2 h (37 °C) before initiating phosphorylation reactions by addition of substrate. CI, confident intervals. Data represent average values ± s.e.m., n = 3. c, Gel-ABPP data showing concentration-dependent partial blockade of YZ-01-A engagement of N112-CCNE1 by I-125A and I-198, but not dinaciclib. Assays were performed in lysates from transfected HEK293T cells co-expressing NLuc-N112C-CCNE1 or NLuc-WT-CCNE1 and CDK2 that were treated with the indicated concentration ranges of compounds followed by YZ-01-A (10 μM, 90 min), anti-FLAG immunoprecipitation, and processing for gel-ABPP. d, Quantification of gel-ABPP data shown in c. Data represent average values ± s.e.m., n = 3. e, NanoBRET-ABPP data showing concentration-dependent partial blockade of YZ-01-A engagement of N112-CCNE1 by I-125A and I-198. Samples were treated with compounds as described in c prior to NanoBRET measurement. Data represent average values ± s.e.m., n = 3. f, Quantification of gel- and NanoBRET-ABPP data showing that dinaciclib does not substantially affect YZ-01-A engagement of N112C-CCNE1. Data represent average values ± s.e.m., n = 3.