Cell Cycle and Viability Analyses

Western blotting, cell viability assays, and cell cycle distribution analysis were performed according to our previous study [27 (link)]. Antibodies against CENPI (1: 2000, catalog no: ab118796) and MCM4 (1 : 2000, catalog no: ab4459) were purchased from Abcam (Cambridge, MA, USA). An antibody against KNTC1 (1 : 200, catalog no: sc-81853) was purchased from Santa Cruz (Santa Cruz, CA, USA). An antibody against GAPDH (1 : 1000, catalog no: 5174S) was purchased from Cell Signaling Technology (Beverly, MA, USA). HRP-conjugated secondary antibodies derived from rabbits and mice were also purchased from Cell Signaling Technology [anti-rabbit IgG (1 : 5000, catalog no: 7074S) and anti-mouse IgG (1 : 5000, catalog no: 7076S)]. Cell viability was measured using a Cell Counting Kit-8 (catalog no: C008-3, 7seabiotech, Shanghai, China) following the manufacturer's instructions. Cell cycle distribution was measured via flow cytometry with PI/RNase staining buffer (catalog no: 550825, BD Biosciences).