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Figure 3 | Molecular Cancer

Figure 3

From: Specific knockdown of uPA/uPAR attenuates invasion in glioblastoma cells and xenografts by inhibition of cleavage and trafficking of Notch -1 receptor

Figure 3

uPA positively regulates Notch in glioblastoma cells. (A) U251 MG, 5310 and 4910 cells were transfected with pSV (scrambled Vector) or with siRNA against Notch 1 and analyzed for uPA and Notch 1 expression by western blotting. Blots were probed with GAPDH antibody to analyze for equal loading of proteins. (B) Fibrin zymography of U251 MG, 5310 and 4910 cells transfected with pSV (scrambled Vector) and siNotch 1 was performed to analyze for uPA activity. (C) & (D) Quantitative expression of relative uPA expression and uPA activity in pSV and siNotch-transfected cells. (E) Exogenous addition of purified uPA activates Notch 1 receptor in uPAR antisense cells and in U251 MG cells. Whole cell lysates prepared from uPAR antisense and U251 MG cells, which were treated with purified uPA (250 ng), ruPAR (50 ng) and in combination, were checked for Notch 1 expression by Western blotting analysis. Blots were probed with GAPDH antibody to analyze for equal loading of lysates. (F) Whole cell lysates prepared from uPAR antisense cells which were left untreated, treated with purified uPA, transfected with shRNA against Delta and Jagged (Notch 1 ligands) with and without stimulation with purified uPA and probed for Notch 1, Delta and Jagged by Western blotting. Blots were probed with GAPDH antibody to analyze for equal loading of lysates. (G) Quantitative analysis of relative Notch 1 expression represented in (E) and (H) Quantitative analysis of relative Notch 1 expression represented in (F). Values are mean ± S.D. of three independent experiments in comparison with the controls (* P < 0/01, **P < 0.001).

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Molecular Cancer

ISSN: 1476-4598