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. Author manuscript; available in PMC: 2010 Oct 7.
Published in final edited form as: Clin Cancer Res. 2009 Jan 15;15(2):452–459. doi: 10.1158/1078-0432.CCR-08-1631

Figure 5. Effect of Notch1 knockdown on invasion, mRNA expression of uPA, and MMP9 activity in 22Rν1 cells.

Figure 5

(A) Protein level of Notch1 and cleaved Notch1 in control, scrambled siRNA and Notch1 siRNA transfected 22Rν1 cells as determined by Western blot analysis. (B) The photomicrograph shows the number of migratory cells transfected with varying concentrations of Notch1 specific siRNA and scrambled siRNA, Lower panel showing quantification of migratory cells in scrambled and Notch1 specific siRNA transfected cells. The experiment was performed in triplicate and the value obtained from scrambled siRNA transfected cells was set as 100% migration. (C) Semiquantitative RT-PCR analysis shows mRNA expression of uPA in cells transfected with increasing concentration of Notch1 siRNA. Scrambled siRNA was used as control in parallel. Lower panel showing GAPDH as an internal control. (D) Effect of Notch1 knockdown on the promoter activity of MMP9 gene expression in PC3 cells. 2×106 22Rν1 cells cotransfected with either scrambled or Notch1 specific siRNA along with 1 μg of pGL3 or 1μg MMP9 luciferase reporter plasmid and 50 ng of renilla luciferase reporter plasmid as an internal control as described in material and method section. The experiment was performed in quadruplet and the values are showing mean ± SD. Asterisk (*) represents p<0.01 as significant.