Figure 3D illustrates a marked decrease in the cell population in G1 phase while the cell population in G2/M arrest was significantly increased in siMPS1-treated KKU-055 and KKU-213A cell lines. Moreover, significant increases in the sub G1 population (apoptotic cells) were found in siMPS1-treated CCA cell lines at 72 and 96 h (Figure 3E). Western blot analysis confirmed that decreasing of Cyclin B1 (G2/M arrest), increasing of Bax pro-apoptotic and decreasing of Mcl-1 anti-apoptotic markers (apoptosis) after MPS1 knockdown in CCA cell lines (Supplementary Figure S2). This evidence concerns the gene TTK and cholangiocarcinoma.