Fig. 4: Overexpression of LNC CRYBG3 induces aneuploidy in immortalized lung epithelial cells aneuploidy.

A Demonstration of the role of BUB3 and other factors in spindle assembly checkpoint. B Effect of LNC CRYBG3 on cell cycle. Colcemid was used to synchronize BEAS-2B cells at M phase and then release the cells to various cell cycle phases. The protein makers of different phase of cell cycle, Cyclin D1 for G0/G1 phase, phosphorylated H3 (p-H3) for M phase, and Cyclin B1 G2/M phase, was detected by western blot analysis. C The relative protein level of Securin which was detected by western blot analysis. Colcemid were used to synchronize cell cycle of BEAS-2B cells at M phase in normal and LNC CRYBG3 knockdown cell lines. D The cell cycle of A549 cells transfected with negative control RNAs (NC) or LNC CRYBG3 was monitored after cells were released to various phases from synchronized M phase. The proportion of aneuploid cells was assayed by flow cytometry. All experiments were repeated at least three times independently. The quantitative data were presented as mean ± SEM. E A confocal time-lapse was used to monitor the cell mitosis of BEAS-2B cells overexpressing control RNA (NC) or LNC CRYBG3. F, G Chromosome stability were visualized and quantified by karyotype analysis after BEAS-2B cells were transfected with LNC CRYBG3 and/or Bub3 shRNA. H, I, J The proportion of aneuploidy in human peripheral blood lymphocyte after irradiation (2.5 Gy X-rays). K, L The expression of LNC CRYBG3 and Bub3 in human peripheral blood lymphocyte after irradiation (2.5 Gy X-rays), measured by qRT-PCR The animal protocol was approved by the Research Ethics Committee of Soochow Medical University, China and the written informed consent was obtained from all subjects. *p < 0.05; **p < 0.01; **p < 0.001.