Expression profiling and functional study of NCL in breast and colorectal cancer. A, A significant correlation between tumor occurrence breast carcinoma or colorectal adenocarcinoma patients and the expression levels of NCL mRNA was observed in the Curtis and Alon datasets, which were obtained through the Oncomine dataset repository (www.oncomine.org). B, Kaplan-Meier survival analyses were conducted for patients with breast and colorectal cancer based on NCL expression in two independent cohorts (Bertucci and Sveen). C, D, Immunohistochemistry assay (IHC) for NCL in normal breast and breast cancer tissues (C) and normal colon and colon cancer tissues (D). Scale bar, 100 µm. E-H, Cancer and normal cells were treated with the indicated amounts of anti-NCL antibody or IgG in serum-free media for 24 h. Cell viability of MCF-10A (E), MDA-MB-231 (F), CCD-18Co (G), and HCT-116 (H) was measured by WST assay. I, Apoptotic cells were visualized by staining Annexin V+ cells. Cells were neutralized with the anti-NCL antibody and incubated in serum-free media for 24 h. Apoptotic cells were measured by FACS analysis. Bar graphs represent the mean ± SD, and statistical analyses were performed by one-way ANOVA with Dunnett's multiple comparison. *, **, and *** indicate P < 0.05, P < 0.01, and P < 0.001, respectively.