Zhang et al. Journal of Experimental & Clinical Cancer Research 2010, 29:88 http://www.jeccr.com/content/29/1/88 Page 7 of 10 Figure 2 A Expression of E-Cadher mRNA in QBC939, SK-Ch-1, FRH 0201 cells. In vitro cleavage effect of different ribozymes on E-Cadherin mRNA and Slug mRNA. The reaction product of in vitro ribozyme cleavage was analyzed by absolute real-time quantitative PCR. The amplification plots and standard curve were obtained with the in vitro transcript from E-Cadherin. Serial 10-fold dilutions with 9 × 108 to 9 × 10- 2 pg per reaction well were made in EASY Dilution (Takara). Amplification was repeated three times for each dilution. It showed the cell line FRH 0201 had the highest expression of E-Cadherin m RNA and the lowest expression of Slug mRNA. B Evaluation of transfection efficiencies. It showed the transfection efficiency was 43.6% 48 h after Slug transfection. C E-cadherin in Slug transfected and mock-transfected FRH 0201 cells. In vitro cleavage effect of different ribozymes on E-Cadherin mRNA. The reaction product of in vitro ribozyme cleavage was analyzed by absolute real-time quantitative PCR. The amplification plots and standard curve were obtained with the in vitro transcript from E-Cadherin. Serial 10-fold dilutions with 9 × 108 to 9 × 10-2 pg per reaction well were made in EASY Dilution (Takara). Amplification was repeated three times for each dilution. It showed Slug overexpression repressed E-cadherin expression in FRH 0201. Figure 3 A Expression of E-cadherin in QBC939 cells. The reaction product of in vitro ribozyme cleavage was analyzed by absolute real-time quantitative PCR. The amplification plots and standard curve were obtained with the in vitro transcript from E-Cadherin. Serial 10-fold dilutions with 9 × 108 to 9 × 10-2 pg per reaction well were made in EASY Dilution (Takara). Amplification was repeated three times for each dilution. It showed Slug inhibition increased E-cadherin expression in QBC939 cells. B Evaluation of transfection efficiencies. It showed the transfection efficiency was 31.4% 48 h after siRNA-Slug transfection.