Fig. 5: ADAR knockout in MEC1 cells sensitizes toward in vitro treatment.

a Schematic representation of ADAR exon 2 and DNA/protein sequence of the CRISPR/Cas9 target site (protospacer adjacent motif is underlined) for the two ADAR isoforms p110 and p150. Sanger sequence of the target site from MEC1 ADAR-knockout cells is shown below (Y = C or T; M = A or C; R = G or A). b A > I editing of FLNB and BLCAP in MEC1 and MEC1 ADAR knockout (MEC1-KO) cells. c Heat map of editing frequencies of 19 recurrent A > I editing sites in MEC1 and MEC1 ADAR knockout cells. d Unique Alu editing sites and Alu editing index (AEI) for MEC1 and MEC1 ADAR knockout cells. e Representative viability stains (measured by flow cytometry and 7AAD/AnnexinV) and dot plot from n = 4 independent experiments (left graph) and longitudinal cell counts (right graph, n = 3) from MEC1 and MEC1 ADAR knockout cells. f Representative cell cycle stains of MEC1 and MEC1 ADAR knockout cells and statistics from n = 3 independent experiments (mean ± SD). g Heat map of differentially expressed genes in MEC1 versus MEC1 ADAR knockout cells. h MEC1 and MEC1 ADAR knockout cells were treated with different doses of indicated drugs in vitro for 72 h followed by viability measurements using XTT assays (Flu: fludarabine; Ibr: ibrutinib; Ven: venetoclax). Viability of controls (DMSO treated cells) were set to 100%. Significances calculated using unpaired t test (n values indicate independent experiments; horizontal lines in dot plots show mean values); *p = 0.01; **p < 0.01.