Fig 2: RAE1 induces chemoresistance. A, Apoptosis assays using RAE1-overexpressing cells and control cells. Cells were cultured in paclitaxel (upper) or actinomycin D (lower) for 24 h with the indicated final concentration. B, Quantification of cell distribution in apoptosis in indicated cells (Annexin V-FITC+). *P < .05; **P < .01; ***P < .005. C, Western blot analysis for RAE1 and cleaved PARP in control cells or RAE1-overexpressing cells treated with 2 nmol/L drug (paclitaxel +) or no drug (paclitaxel -). D, Sensitivity of RAE1-overexpressed cells to paclitaxel (left) or actinomycin D (right). Cell viability was measured by MTT assays in RAE1-overexpressing cells and control cells treated with the indicated final concentrations of paclitaxel or actinomycin D for 24 h. ***P < .005. E, MTT assay was used to assess the viability of RAE1-knockdown and control cells treated with the same final concentrations of paclitaxel for 24 h. ***P < .005

Fig 3: RAE1 is a gene that potentially promotes tumor progression. A, DNA copy number variations in chromosomal arms in 617 colorectal cancer (CRC) tissues using data from The Cancer Genome Atlas (TCGA). B, RAE1 mRNA expression in CRC tissues and normal colon tissues in TCGA dataset and our CRC cohort. ***P < .005. C, Correlation between DNA copy number and mRNA expression of RAE1 in TCGA dataset and Cancer Cell Line Encyclopedia (CCLE) dataset. R is the Pearson correlation coefficient. D, Immunohistochemical staining for RAE1 in CRC tissues and normal tissues. N, normal tissue; T, tumor tissue. E, Pie chart of RAE1 mutation distribution in CRC cases of the Catalogue of Somatic Mutations in Cancer (COSMIC) database (n = 2547). F, Kaplan-Meier overall survival curves of CRC patients according to RAE1 mRNA expression in TCGA dataset (n = 597), our CRC cohort (n = 102)

Fig 4: RAE1 expression is associated with cell growth of colorectal carcinoma. A, RAE1 mRNA expression using RT-quantitative PCR (RT-qPCR), and protein expression using western blot analysis in RAE1-knockdown cells and control cells. *P < .05. B, MTT assays using RAE1-knockdown cells. ***P < .005. C, Colony formation assays using RAE1-knockdown cells. ***P < .005. D, RAE1 protein levels in indicated cells. RAE1 protein expression using western blot analysis of the rescued cells. E, MTT assay using indicated cells. *P < .05. F, RAE1 mRNA expression using RT-qPCR and protein expression using western blot analysis in RAE1-overexpressing cells and control cells. ***P < .005. G, MTT assay using RAE1-overexpressing cells. ***P < .005. H, Colony formation assay using RAE1-overexpressing cells. *P < .05 I, In vivo analysis using a xenograft model. Tumor size in RAE1-knockdown cells and control cells. *P < .05; **P < .01; ***P < .005. J, Immunohistochemical staining for RAE1 in tumor tissues from control and RAE1-knockdown cells. Original magnification, ×400

Fig 5: RAE1 is associated with spindle bipolarity in mitosis. A, Immunofluorescence photographs of CACO2 cells stained with anti–a-tubulin (green) and anti–pericentrin (red) antibodies. Nuclei are stained with DAPI (blue). B, Quantification of mitotic-defective cells with multipolar spindles in RAE1-knockdown cells. *P < .05; **P < .01. C, Immunofluorescence photographs of control cells (left) or RAE1-overexpressing cells (right) treated with 2 nmol/L paclitaxel before staining. D, Quantification of mitotic-defective cells with multipolar spindles in indicated cells. **P < .01