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. 2011 Apr 1;12(5):470–476. doi: 10.1038/embor.2011.39

Figure 4.

Figure 4

Brain endothelial cell secretome can restore mTOR activation in glioblastoma stem-like cells. (A) GSCs were treated with human brain endothelial cell-conditioned medium (EC-CM) for 72 h and compared with control medium (Ctl) and 293T-CM. Protein extracts were tested by western blot with the indicated antibodies. (B,C) GSCs were treated with EC-CM alone or in the presence of rapamycin (RP 50 nM), PP242 (1 μM) and PI103 (10 μM) for 24 h. (D,E) GSCs were transfected with siRNA against mTor, Rictor and Raptor or a control siRNA (sic) for 72 h. (B,D) The number of secondary neurospheres per field of view (FOV) was counted. (C,E) Cell viability was analysed by MTT and normalized to optical density (OD) obtained with DMSO and sic, respectively. All graphs represent mean+s.e.m. of three independent experiments. Analysis of variance test: ***P<0.001, **P<0.01, *P<0.05. DAPI, 4,6-diamidino-2-phenylindole; GSC, glioblastoma stem-like cell; mTOR, mammalian target of rapamycin; siRNA, small-interfering RNA.