Supplementary Figure 12: Base editing induced mutational activation of CTNNB1 enables outgrowth following Tankyrase and MEK inhibition
From: Optimized base editors enable efficient editing in cells, organoids and mice
a. Schematic overview of the fluorescence-based competitive proliferation assay. Parental cells are shown in gray, transduced cells (tdTomato+) are in red, and cells bearing the target editing are highlighted in blue. Neutral competition keeps both tdTomato+ and tdTomato- cell proportions constant, whereas positive or negative selection causes the tdTomato+ population to increase or decrease, respectively. b. BE3, RA, 2X, and FNLS-expressing DLD1 cells were transduced with CTNNB1.S45 sgRNAs and treated with DMSO (left) or XAV939 1uM + Trametinib 10nM (right). Graph shows the number of tdTomato+ cells relative to the start of the assay. Bars represents measurements every 5 days (0, 5, 10, 15). Graphs show mean values. Error bars represent s.e.m., n = 3 biologically independent experiments; asterisks (*) indicate a significant difference (p<0.05) between groups, using an ANOVA with Tukey’s correction for multiple testing. c. Same as in b. but using FANCF.S1 (control) sgRNA. Note the neutral impact on relative proliferation in all the conditions, in contrast to CTNNB1.S45.
