FIGURE 7 from Induction of Viral Mimicry Upon Loss of DHX9 and ADAR1 in Breast Cancer Cells
Year: 2024
Type: other
Abstract: Rescue of PKR activation by DHX9 mutants in MCF-7. A, Representative immunoblot showing the phenotype of ADAR1 and DHX9 knockdown with DHX9, DHX9K417R or dsRBD-EGFP overexpression in MCF-7. Immunoblots for other replicates and uncropped blots can be found in Source Data Figures. B, Fold change of PKR phosphorylation at Thr-446 upon ADAR1 and DHX9 knockdown with DHX9, DHX9K417R or dsRBD-EGFP overexpression in MCF-7, quantified from immunoblots in A. Quantification of protein expression for other proteins of interest can be found in Supplementary Fig. S9B–S9G. C, Representative foci formation phenotype of ADAR1 and DHX9 knockdown with DHX9, DHX9K417R or dsRBD-EGFP overexpression in MCF-7. Quantification of relative foci area is shown in D. E, RNA expression of representative genes from upregulated pathways in Fig. 5 as determined by qRT-PCR for the MCF-7 rescue experiment. Statistical analysis for each pathway is shown in Supplementary Fig. S9H–S9J. F, Analysis of rRNA integrity upon knockdown of ADAR1 and DHX9 in MCF-7 with overexpression of EGFP or dsRBD-EGFP. Arrows indicate canonical RNase L cleavage products (64). Timepoints for collecting protein lysates, isolating RNA, and foci formation are the same as described in Fig. 4 for MCF-7. Bars represent the average of at least three replicates, error bars are ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001. P values determined by Dunnett test.
Language: English
Topic: RNA regulation and disease
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4393949966