Fig. 1: PACT restricts aberrant activation of PKR, thereby maintaining cellular homeostasis.
From: PACT prevents aberrant activation of PKR by endogenous dsRNA without sequestration
A ATP bioluminescence assay showing cell viability in PACT KO-sensitive (HCC1806, MDA-MB-157, NCI-H727, NCI-H2286, SW1271) and -insensitive (ZR-75-1, HCC1428, NCI-H1437, NCI-H2023, NCI-H1650) cell lines after control gene-KO (ctrl) or PACT-KO using 2 different guide RNAs. The control (ctrl) genes are AAVS1 (sg1) and Chr2.2 (sg2). Values represent means ( ± SD) of 3 biological repeats. P values were based on one-way ANOVA test. B Crystal violet staining assay showing cell viability in 3 PACT KO-sensitive cells (HCC1806, NCI-H727, and NCI-H2286) after control gene-KO vs. PACT-KO. C Crystal violet staining assay with HCC1806 cells after control gene-KO and PACT-KO complemented with increasing amounts of wild-type PACT. PACT used here was engineered to be resistant to PACT sgRNA 1 targeting. D Western blot analysis showing levels of PKR and eIF2α phosphorylation in PACT KO-sensitive (top panel) and -insensitive (bottom panel) cells after control gene-KO or PACT-KO. Vinculin was used as a loading control. The experiment was repeated 3 times independently with similar results. E ATP bioluminescence assay in HCC1806 cells in PKR-sufficient and PKR-KO cells in control gene-KO and PACT-KO backgrounds. Values represent means ( ± SD) of 3 biological repeats. P values were based on two-way ANOVA test. F A representative crystal violet staining for samples in E. G ATP bioluminescence assay in HCC1806 cells in control gene-KO and PACT-KO treated with indicated concentrations of ISRIB. Values represent means ( ± SD) from 4 technical replicates across two independent experiments (n = 2). P values were based on two-tailed student’s t-test. ****p = <0.0001, ***p = <0.001, **p = <0.01, *p = <0.05, not significant (ns) is for p > 0.05. Source data and exact P values are provided as a Source Data file.
