Journal: Cell reports

Article Title: XRN1 deletion induces PKR-dependent cell lethality in interferon-activated cancer cells

doi: 10.1016/j.celrep.2023.113600

Figure Lengend Snippet: (A) Correlation of expression levels of individual genes with XRN1 genetic dependency based on CRISPR-Cas9-mediated gene essentiality screens. Each dot represents one gene, and the top ten gene expression correlates are labeled in red. Pearson correlations and corresponding p adj values were computed for each feature in the Cancer Dependency Map Public 22Q4 dataset using all cancer cell lines. (B) Representative immunoblots from two independent biological replicates showing phospho-STAT1, total STAT1, total PKR, MDA5, and β-actin protein levels in a panel of XRN1 KO-sensitive cancer cell lines treated with either DMSO control or ruxolitinib (1 μM) for 24 h. Molecular weight (MW) markers are shown in kDa. (C, E, and G) Representative immunoblots showing XRN1, phospho-STAT1, total STAT1, phospho-PKR, total PKR, and β-actin protein levels in control and XRN1 -deleted NCI-H1650 (C), HCC1438 (E), and SW900 (G) cells treated with either DMSO control or ruxolitinib (1 μM). MW markers are shown in kDa. Three independent biological replicates were performed for each cell line. (D, F, and H) Cell viability was assessed by either ATP bioluminescence (left panels) or crystal violet staining (right panels) after CRISPR-Cas9 targeting of control loci or XRN1 in NCI-H1650 (D), HCC1438 (F), and SW900 (H) cells treated with DMSO control or ruxolitinib (1 μM). ATP bioluminescence values were normalized to the control sg1 sample within each cell line. Each dot represents the average of three technical replicates from one of three independent biological replicates in (D), (F), and (H). Error bars represent standard deviation from the mean. *p < 0.05 and ***p < 0.001, as calculated by repeated measures two-way ANOVA. Crystal violet images are representative of three independent biological replicates. See also .

Article Snippet: Rabbit polyclonal anti-total STAT1 , Cell Signaling Technology , Cat# 9172; RRID: AB_2198300.

Techniques: Expressing, CRISPR, Gene Expression, Labeling, Western Blot, Control, Molecular Weight, Staining, Standard Deviation

Journal: Cell reports

Article Title: XRN1 deletion induces PKR-dependent cell lethality in interferon-activated cancer cells

doi: 10.1016/j.celrep.2023.113600

Figure Lengend Snippet: (A) Representative immunoblots showing XRN1, phospho-STAT1, total STAT1, MDA5, phospho-PKR, total PKR, and β-actin protein levels in control or XRN1 KO A549 (left) or NCI-H1299 (right) cells after 24 h of treatment with vehicle control (sterile water) or interferon-β (10 ng/mL). Molecular weight (MW) markers are shown in kDa. (B) Cell viability was assessed by ATP bioluminescence in control or XRN1 KO A549 (left) or NCI-H1299 (right) cells 5 days after treatment with vehicle control (sterile water) or the indicated concentrations of interferon-β. Each dot represents the average of three technical replicates from one independent experiment. (C) Representative immunoblots showing XRN1, phospho-PKR, total PKR, and β-actin protein levels in control, XRN1 single KO, PKR single KO, or XRN1/PKR double KO (DKO) A549 (left) or NCI-H1299 (right) cells after 24 h of treatment with vehicle control (sterile water) or interferon-β (10 ng/mL). MW markers are shown in kDa. (D) Cell viability was assessed by ATP bioluminescence in control, XRN1 single KO, PKR single KO, or XRN1/PKR double KO (DKO) A549 (left) or NCI-H1299 (right) cells 5 days after treatment with vehicle control (sterile water) or the indicated concentrations of interferon-β. Each dot represents the average of three technical replicates from one independent experiment. Three independent biological replicates were performed for each cell line in (A)–(D). ATP bioluminescence values were normalized to the vehicle control sample for each isogenic cell line in (B) and (D). Error bars represent standard deviation from the mean. See also .

Article Snippet: Rabbit polyclonal anti-total STAT1 , Cell Signaling Technology , Cat# 9172; RRID: AB_2198300.

Techniques: Western Blot, Control, Sterility, Molecular Weight, Standard Deviation

Journal: Cell reports

Article Title: XRN1 deletion induces PKR-dependent cell lethality in interferon-activated cancer cells

doi: 10.1016/j.celrep.2023.113600

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-total STAT1 , Cell Signaling Technology , Cat# 9172; RRID: AB_2198300.

Techniques: Recombinant, Protease Inhibitor, Western Blot, Stripping, Bicinchoninic Acid Protein Assay, Transfection, Cell Viability Assay, Gene Expression, Control, CRISPR, Gene Knockout, Clone Assay, Plasmid Preparation, Expressing, Software