Journal: The Journal of Clinical Investigation
Article Title: High PRMT5 levels, maintained by KEAP1 inhibition, drive chemoresistance in high-grade serous ovarian cancer
doi: 10.1172/JCI184283
Figure Lengend Snippet: ( A ) Schematic displays TMAs used for PRMT5 staining. Figure was created with BioRender.com. ( B ) Representative IHC images show QuPath annotation of tumor stroma and epithelial cells (left), and a violin plot shows PRMT5 staining intensity in stroma versus tumor epithelial cells (right). P value was determined by Wilcoxon rank-sum test. Original magnification: ×5. ( C ) Representative IHC images show PRMT5 staining in primary and recurrent tumors. Original magnification: ×5. ( D ) The density plot displays H-scores for PRMT5 staining in primary tumors (left), and the dot plot shows PRMT5 levels (H-score) between primary and recurrent tumors for the primary tumors with low PRMT5 staining (right). P values were quantified by 2-tailed, paired Student’s t test. ( E ) Western blots show PRMT5 and SDMA levels in chemo-naive and -resistant isogenic cell line pairs. Noncontiguous lanes on different blots have been separated by thin dashed lines. ( F ) Western blots show PRMT5 expression after different doses of Dox induction (72 hours) in chemo-naive OVCAR4 cells. Carboplatin-resistant OVCAR4 cells were used to determine Dox levels for overexpression. ( G ) Line plots show the relative apoptosis rate of OVCAR4 cells treated with the indicated doses of Dox. Cells were subjected to 40 μM carboplatin, and apoptosis (caspase 3/7 activity) was monitored by the IncuCyte live-cell imaging platform for 72 hours. Data are shown as the mean ± SEM ( n = 3). P values were quantified by 1-way ANOVA with Dunnett’s multiple-comparison test. ( H ) Western blots show PRMT5 levels in OVCAR4 cells expressing sgRNAs targeting PRMT5 promoter. ( I ) Line plots show the relative apoptosis rates of OVCAR4 cells expressing the indicated sgRNAs. Cells were treated with 20 μM carboplatin, and apoptosis was monitored over 72 hours using the IncuCyte live-cell imaging platform. Data are shown as the mean ± SEM ( n = 3). P values were determined by 1-way ANOVA with Dunnett’s multiple-comparison test.
Article Snippet: The following day, treatments were administered using a drug of interest mixed with a 1:1,000 dilution of caspase 3/7 dye (10403, Biotum) and a 1:2,000 dilution of SiRDNA nuclei stain (CY-SC007, Spirochrome).
Techniques: Staining, Western Blot, Expressing, Over Expression, Activity Assay, Live Cell Imaging, Comparison
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