Fig 2: NUSAP1 expression and prognosis value in ovarian epithelial malignancies. a Representative images of NUSAP1 expression in ovarian tissue: epithelial ovarian cancer, epithelial ovarian borderline tumor, benign ovarian epithelial tumor, and normal ovarian tissue. Scale bar: 100 µm (upper picture) and 50 µm (lower picture). The lower picture is an enlarged view of the blue box in the upper picture. b Correlation between NUSAP1 expression, FIGO stage, residual tumor size, and survival prognosis in patients with epithelial ovarian malignancies

Fig 3: NUSAP1 downregulation induced cell cycle arrest and apoptosis in BLCA cells. (A). GSEA suggests that NUSAP1 is closely related to the G2/M cell cycle phase. (B and C). Cell cycle analysis via flow cytometry. **P<0.01 vs NC. (D). Western blotting analysis of CDK1 and cyclin B1 protein expression. (E and F). Apoptosis was analyzed by flow cytometry. **P<0.01 vs NC (5637), *P<0.05 vs NC (T24). (G). Western blotting analysis detected the protein expression levels of Bax, Bcl-2 and cleaved-caspase3.

Fig 4: NUSAP1 regulated the epithelial-mesenchymal transition of BLCA via the TGF-ß signaling pathway. (A). GSEA revealed that NUSAP1 expression was positively correlated with the TGFBR complex and the Smad2/3 pathway in BLCA. (B). Gene correlation analysis of NUSAP1 and TGFBR1 obtained from Gene Expression Profiling Interactive Analysis (GEPIA, http://gepia.cancer-pku.cn/). ***P<0.001. (C). Real-time PCR was performed to detect the relationship between NUSAP1 and TGFBR1. **P<0.01, vs NC (5637) or NC (T24). (D). Western blotting analysis of the protein expression levels of TGFBR1, Smad2/3, p- Smad2/3, E-cadherin, and vimentin. (E and F). Wound healing assays were performed to detect the migration of cells (100× magnification) *P<0.05, **P<0.01. (G and H). Transwell assays were performed to evaluate the migration and invasion of cells (100× magnification). *P<0.05, **P<0.01, ***P<0.001. (I). Western blotting analysis of the protein expression levels of Smad2/3, p- Smad2/3, E-cadherin, and vimentin.

Fig 5: NUSAP1 overexpression significantly increases metastases in a PC-3-luc2 xenograft modelMice bearing PC-3-luc2 flank tumors overexpressing NUSAP1 or EGFP were assessed for metastases to the lung, axillary lymph nodes (right and left), femoral bone marrow (right), liver, or spleen by measuring expression levels of human GAPDH relative to universal GAPDH by RT-qPCR. Measurements of metastases in A. all six tissue sites combined and the B. lung, C. right axillary lymph node, D. left axillary lymph node, E. right femoral bone marrow, F. liver, and G. spleen. All horizontal lines represent the median. All P-values were calculated using the Mann-Whitney U test.