Fig 2: Downregulation of ATP1A1 expression by small interfering RNA (siRNA) decreases ESCC cell invasion, migration and colony formation in vitroA. Endogenous ATP1A1 RNA expression in the CE81T and CE81T-4 cells; B. Down-regulated ATP1A1 mRNA levels by ATP1A1 siRNA; C. Down-regulated ATP1A1 protein expression by ATP1A1 siRNA; D. Inhibition of cell proliferation in CE81T and CE81T-4 cells by ATP1A1 siRNA; E. Decreased colony formation in CE81T and CE81T-4 cells by ATP1A1 siRNA. F. Decreased cell migration ability using wound-healing assay in CE81T and CE81T-4 cells by ATP1A1 siRNA (Original magnification: ×200). G. Decreased invasion ability in CE81T and CE81T-4 cells by ATP1A1 siRNA. Data are quantified using Image J software and error bars represent s.d. of means for experiments in triplicate (***P value<0.005,**P value<0.001, *P value<0.05, respectively, Student's t-test).

Fig 3: Identification of up-regulation candidate genes from Rat One array and selection of candidate genes for clinical applicationA. The flow chart for identifying candidate genes (Control: DMSO treated only; Normal: normal part from NMBA and arecoline treated rats; Tumor: tumor part from NMBA and arecoline treated rats); B. ATP1A1 RNA levels in tumor and adjacent normal tissues from 14 ESCC patients by real-time PCR ((Calculated by 1000 × 2–(ATP1A1_AverageCT – 18S_AverageCT); C. The distribution of relative ATP1A1 RNA expression levels (tumor vs. adjacent normal parts) by boxplot and dotplot (Minimum: 0.24, the first quartile: 0.99, median: 3.49; the third quartile: 4.36, and maximum: 34.40). Wilcoxon signed rank test, p = 0.0052.