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. 2016 Oct 15;30(20):2259–2271. doi: 10.1101/gad.287474.116

Figure 7.

Figure 7.

Depletion and inhibition of USP10 impairs the growth of aneuploid human cells. (A) RPE-1 cells were treated with siRNA targeting USP10 or control siRNA. Twenty-four hours later, cells were treated with either MAD2 siRNA or control siRNA or reversine or solvent. Three days after Mad2 siRNA or reversine treatment, cells were harvested, and the number of cells was determined. n = 3. Mean ± SD shown. P < 0.05, Student's t-test. (B) RPE-1 cells were treated with siRNA targeting Mad2 or control siRNA. Twenty-four hours later, cells were treated with spautin-1 or solvent only. Three days after spautin treatment, cells were harvested, and the number of cells was determined. n = 3. Mean ± SD shown. P < 0.01, Student's t-test. (C) RPE-1 cells were treated with reversine or solvent to induce chromosome missegregation. Twenty-four hours later, the drug was washed out, and cells were treated with spautin-1 or solvent. Three days after spautin treatment, cells were harvested, and the number of cells was determined. n = 3. Mean ± SD shown, P = 0.0015, Student's t-test. (D) MEFs either euploid or trisomic for chromosome 13 or 16 were treated with spautin-1 or solvent only. Three days after treatment, cells were harvested, and the number of cells was determined. n = 3. Mean ± SD shown. P < 0.05, Student's t-test. (E) RPE-1 cells were treated with spautin-1 or solvent. Twenty-four hours later, cells were treated with reversine or solvent to induce chromosome missegregation. Three days after reversine treatment, cells were treated with Bafilomycin A1 (BafA1) or solvent only and harvested 6 h later. LC3-I and LC3-II levels were analyzed by Western blot analysis. GAPDH was used as a loading control. (F) Quantification of LC3-II levels in E were normalized to the untreated control (lane 1). (G) Levels of USP10 gene expression in matched normal and cancer cells were plotted. Data are from the Broad Institute GDAC Firehose. (BRCA) Breast-invasive carcinoma; (COADREAD) colorectal adenocarcinoma; (HNSC) head and neck squamous cell carcinoma; (KIPAN) pan-kidney cohort; (LIHC) liver hepatocellular carcinoma; (LUAD) lung adenocarcinoma; (LUSC) lung squamous cell carcinoma; (PRAD) prostate adenocarcinoma; (STAD) stomach adenocarcinoma. (*) P < 0.0001.