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Figure 2 | Molecular Cancer

Figure 2

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

Figure 2

Transcriptional inactivation of uPA and/or uPAR by shRNA inhibits cleavage of Notch 1 receptor, Notch 1 gene expression in U251 MG, 4910 and 5310 xenograft cells. (A) Whole cell lysates were prepared from U251 MG, 4910 and 5310 cells which were left untransfected, or transfected with pSV, puPA, puPAR and pU2 to evaluate the protein levels of activated Notch 1 and for Notch 1 cleavage at the Val 1744 position. Blots were probed with GAPDH antibody to analyze for equal loading of proteins. (B) & (C) Quantitative analysis of relative Notch 1 expression and cleaved Notch expression in U251 MG, 5310 and 4910 cells. (D) Immunofluorescence analysis of U251MG cells left untransfected or transfected with pSV, puPA, puPAR and pU2 and stained for NICD (Notch intracellular domain) and images taken with Olympus Fluoview microscope. (E) RT-PCR analysis of Notch 1 mRNA expression in (a) U251 MG, 5310 and 4910 cells which were left untransfected or transfected with pSV, puPA, puPAR and pU2. GAPDH was used as an internal control. (F) Quantitative representation of Notch 1 mRNA levels in U251MG, 5310 and 4910 cells. Values are mean ± S.D. of three independent experiments (* P < 0.01, **P < 0.001).

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Molecular Cancer

ISSN: 1476-4598