Fig. 1
Structural, pharmacological, and biochemical characterization of JNJ-BJ. a Chemical structure of JNJ-BJ. The compound is the first eluted enantiomer of a 3-ethylquinolinone bearing a chiral center linked to the C-7 carbon atom ((R or S)-3-[2-cyano-2-(3-ethyl-2-oxo-1H-quinolin-7-yl)propyl]benzonitrile). b Tankyrase (TNKS) autoribosylation assay. A recombinant His-tagged human TNKS2 poly(adenosine diphosphate)-ribose polymerase domain was produced in a baculovirus/insect cell expression system. The purified protein was bound to a 384-well Ni2+-coated Flashplate and incubated for 120 min with NADH3/NAD in the presence of increasing concentration of JNJ-BJ. The radioactive signal was measured using a scintillation reader. c Assessment of β-catenin/TCF transcriptional activity through a TOPflash reporter assay in adenomatous polyposis coli-mutant DLD1 colorectal cancer cells. Cells were treated with TNKS/2 inhibitors (10 μM) for 24 h. Results are expressed as TOP/FOP ratio and represent the average (lines) ± range of two independent experiments (diamonds), each performed in technical quadruplicate. d Expression of AXIN2 and LGR5 transcripts in DLD1 cells treated with TNKS/2 inhibitors (10 μM) for 24 h. Results are the average (lines) of two independent experiments (diamonds), each performed in technical triplicate. Raw data for panels c and d are shown in Additional file 2. DMSO dimethyl sulfoxide, IC 50 half-maximal inhibitory concentrations