Figure 1
Expression of Notch signaling components in glioblastoma cancer cell line and xenograft cell lines. (A) Representative composite of Notch 1, 2, 3 and 4 receptors by immunohistochemistry in glioblastoma human tissue array. (B) Western blot analysis of Notch signaling components in three different cells, namely U251 MG and xenograft cell lines 5310 and 4910. (C) RT-PCR analysis of Notch 1, 2, 3 and 4 in U251 MG and xenograft cell lines, 5310 and 4910. (D) shRNA against uPA, uPAR and U2 inhibits anchorage independent growth of U251 MG and glioblastoma xenograft cell lines in soft agar. (Panels a to e - U251 MG, panels f to j - 5310 and panels k to o - 4910). A total of 1 × 105 cells from each cell line were left either untransfected or transfected with pSV, puPA, puPAR and pU2, and these cells were then plated in soft agar. Representative photographs are shown which were taken at four weeks after the colonies were stained with crystal violet. (E) Quantitative representation of number of anchorage-independent colonies in soft agar colony formation assay before and after transfection with pSV, puPA, puPAR and pU2. Values are mean ± S.D. of three independent experiments, in comparison with controls (* P < 0.01, ** P < 0.001) (F) shRNA against uPA, uPAR and U2 inhibits GBM invasion by a matrigel invasion assay as described previously in Material and Methods.