Fig. 2: ADAR1 silencing is synthetically lethal with BRCA1/2 mutations in multiple model systems in vitro and in vivo. | Nature Communications

Fig. 2: ADAR1 silencing is synthetically lethal with BRCA1/2 mutations in multiple model systems in vitro and in vivo.

From: Autocrine interferon poisoning mediates ADAR1-dependent synthetic lethality in BRCA1/2-mutant cancers

Fig. 2

A Schematic describing the isogenic and non-isogenic cell line models used throughout the study. B, C Clonogenic survival of MEF Brca1-wildtype (WT) and Brca1-mutant (Δ11) cells transfected with a concentration range (nM) of Adar1 siRNA. Violin plots indicate median, lower and upper quartiles; N = 6 values from individual wells, representative of n = 3 biologically-independent experiments, two-way ANOVA post hoc Dunnett’s test. P values, *=0.0402, **=0.0017, ****< 0.0001. D Heatmap showing surviving fractions elicited by ADAR1 suppression in models evaluated in Figs. 1, 2 and Supplementary Fig. 1–3 (blue, SF > 0.8; red, SF < 0.8). E, F Cell survival of HEK293T ADAR1-wildtype (WT) and ADAR1-knockout (KO) cells transfected with a concentration range (nM) of BRCA1 (E) or BRCA2 (F) siRNA. Box-and-whiskers indicate median, lower and upper quartiles, and the min to max range; N = 4 values from individual wells, representative of n = 3 biologically-independent experiments, two-way ANOVA post hoc Dunnett’s test. P values, **[ADAR1-WT, siBRCA2 2 nM]=0.001 (F), ***[ADAR1-KO, siBRCA1 2 nM]=0.0009 (E), ***[ADAR1-KO, siBRCA1, 4 nM]=0.0005 (E), ****<0.0001. G–J Representative images and quantifications of morphological phenotypes (G, I) and acridine orange staining (H, J) in zebrafish embryos subjected to morpholino (MO)-mediated knockdown of brca2 and/or adar1. Dead embryos are indicated with an asterisk (G). White arrows indicate acridine orange-positive cells on images taken within a defined region along the anterior-posterior axis (H); scale bar, 500 µm. Percentages of morphological phenotypes (I) were calculated based on N = 149 [no MO], N = 95 [control MO], N = 109 [brca2 MO], N = 97 [adar1 MO] and N = 174 [brca2/adar1 MOs] values from individual embryos, representative of n = 3 biologically-independent clutches. Violin plots indicate median, lower and upper quartiles; N = 31 [no MO], N = 33 [control MO], N = 25 [brca2 MO], N = 23 [adar1 MO] and N = 24 [brca2/adar1 MOs] values from individual embryos, representative of n = 3 biologically-independent experiments, Kruskal-Wallis test post hoc Dunn’s test. P value, ****<0.0001. siCTRL, non-targeting, negative control siRNA; siPLK1, PLK1-targeting, positive control siRNA. Source data are provided as a Source Data file. Elements of panel A were provided by Servier Medical Art (https://smart.servier.com/), licensed under CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/).

Back to article page