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. Author manuscript; available in PMC: 2012 Jul 8.
Published in final edited form as: Cell. 2011 Jul 8;146(1):53–66. doi: 10.1016/j.cell.2011.06.006

Figure 4. GSC cell cycle flux is supported by NOS2 activity, which modulates gene expression including the cell cycle inhibitor CDA1.

Figure 4

(A) 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was employed to evaluate the effect of NOS2-directed shRNA on S-phase transit in CD133+ glioma cells (GSCs). (B) Microarray analysis demonstrated that NOS2-directed shRNA increased transcript expression of the cell cycle inhibitor, CDA1 (2 xenografts in duplicate). NOS2-dependent suppression of CDA1 in GSCs was validated by (C) qRT-PCR and (D) Western analysis. (E) Viability of GSCs and CD133− cells (non-GSCs) was evaluated after treatment with vector or CDA1 expressing lentivirus. F) Neurosphere formation capacity was evaluated 10 d after single vector or CDA1-overexpressing cells were sorted into wells. (G) The decreased EdU incorporation from NOS2-directed shRNAs was partially blocked by concurrent expression of CDA1-directed shRNA. *, p<0.05; **, p < 0.01; ***, p < 0.001, N.S., not significant. See also Figure S4 and Table S1.