Figure 2.

Six1 knockdown can inhibit clone formation and sensitize cells to 5-Fu treatment. (A) Western blot assay demonstrated that Six1 protein level was decreased with siRNA treatment. β-actin was used as internal control. (B) Six1 transcriptional level was knocked down successfully after siRNA transfection with qRT-PCR detection. (C) IFC staining confirmed Six1 protein was decreased in nuclear after siRNA transfection. (D) The representative picture of clone formation with Six1 silence. (E) The histogram indicated the average number of survival clones in NC and Six1 knock-down groups. (F) Cell viability was measured by CCK-8 assays. The percentage of survival cells reduced more significantly in Six1 knock-down groups when various dose 5-Fu drugs were added. (G) Cell cycle transition was analyzed by flow cytometry. The percentage of each cell cycle phase was not affected by Six1 reduction. Data were shown as mean ± SD (n=3), and *, p < 0.05.