Fig 1: Effects of U50A on mitosis and colony-forming ability of breast cancer cells. Nocodazole (100 nM) was used to treat U50A-overexpressing MCF-7 cells for 14 h and was removed at the indicated time points. Cells were collected and subjected to immunofluorescence. (A) Representative images of immunofluorescence in U50A-overexpressing and control MCF-7 cells at the indicated time points. Yellow arrowheads indicate prometaphase cells, and red arrowheads indicate metaphase/anaphase cells. (B) Quantitated results of prometaphase, metaphase, and anaphase cells from immunofluorescence images. Colony forming ability of breast cancer cells expressing U50A (C) and inhibiting U50A (D) were assayed. (E) Specific shRNAs were used to inhibit CENPE/F, ATRX, and SMC5 in U50A-inhibiting BT-474 cells and colony forming capacity were assayed. Data are represented as the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.