Fig. 3

Cell lethality after ADAR deletion is partially mediated through activation of PKR signaling. a Analysis of differentially expressed genes between ADAR KO-sensitive and KO-insensitive cancer cell lines using gene expression data from CCLE¹⁷. Differentially expressed genes are plotted by −log(q-value) on the y-axis versus log2(fold change) on the x-axis. b Immunoblots showing phosphorylated (Thr-446) and total PKR protein levels 5 days after ADAR deletion by CRISPR-Cas9 for the indicated cell lines (n = 5). β-Actin served as a loading control. c Heat maps showing standardized t-statistics of normalized expression values for the indicated genes (rows) after deletion of GFP (control) or ADAR (columns) in the indicated cancer cell lines (n = 1). Color scales show relative normalized expression between ADAR and GFP KO samples. d Cell viability of control or PKR-deficient HCC366 cells was assessed by crystal violet staining 8 days after GFP or ADAR KO with CRISPR-Cas9. A representative image of crystal violet staining (left) and quantitation of cell viability (right) from two independent biological replicates are shown. Cell viability values were normalized to the GFP sg2 control #2 within each group of isogenic cell lines. e Cell viability of control, PKR-deficient, ADAR1-deficient, or ADAR1/PKR double-deficient A549 cells was assessed by ATP bioluminescence 3 days after treatment with vehicle or IFN-β (10 ng/mL). ATP bioluminescence values were normalized to the vehicle-treated control within each isogenic cell line. Data from two biological replicates are shown. Note: ADAR sg2 was used in this experiment. Error bars represent standard deviation in all graphs