CTNNB1 contributes to the cytotoxic effect of AS1842856. (A-B) 018Z and RS4;11 cell lines were CRISPR/Cas9 edited with tandem CRISPR-RNA (crRNA) targeting CTNNB1. (A) The efficiency of CTNNB1 loss-of-function editing was measured by immunoblotting after 24-hour incubation with AS1842856 at a concentration of 100 nM to stabilize CTNNB1 (n = 2). (B) Dynamic of CTNNB1 inactivating mutations in CRISPR/Cas9-edited RS4;11 and 018Z cell lines. The proportion of inactivating mutations (KO score) was monitored by quantification of Sanger chromatograms of the edited region using the ICE (Inference of CRISPR Edits) CRISPR analysis tool (https://ice.synthego.com, accessed on 13 June 2024; Synthego Corporation, Redwood City, CA). (C-I) CTNNB1-KO decreases the sensitivity of B-ALL to AS1842856 and CHIR-99021. The cytotoxic effect of the inhibitors on the survival of 018Z (C-E) and RS4;11 cell lines (F-I) was analyzed with the help of 6-day MTT assay. In panels E,H-I, the dose-effect data were fitted by nonlinear regression model and IC50 values and AUC values were calculated with the help of GraphPad Prism software, version 10.30 (San Diego, CA). The significance of IC50 or AUC differences was calculated by 2-sided nonpaired Student t test (n = 3).AUC, area under survival curve.
