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. Author manuscript; available in PMC: 2013 Jul 10.
Published in final edited form as: Oncogene. 2012 Jan 16;31(44):4709–4717. doi: 10.1038/onc.2011.622

Figure 1. shRNA screen design: Active Rb expression induces stable cell cycle arrest and senescent phenotype.

Figure 1

(A) T98G glioblastoma cells expressing the murine ecotropic receptor (T98G-EcoR) were infected with empty retrovirus or retrovirus producing a PSM-Rb. Three days post-selection in hygromycin, cells were labeled with BrdU for 16 h, fixed and BrdU incorporation was detected by immunofluorescence. At least 200 cells were counted per condition. Bar graphs represent mean ± S.D. from two independent experiments for PSM-Rb. (B) T98G-EcoR cells treated as in (A) were stained to detect SA-beta-galatosidase activity. (C) The shRNA screen was designed to identify cells resistant to PSM-Rb expression in the shPool populations as compared to the control, shLuc (luciferase). (D) T98G-EcoR cells stably producing shRNA targeting firefly luciferase were infected with empty retrovirus or retrovirus producing PSM-Rb. Either seven days (empty retrovirus) or 19 days (PSM-Rb retrovirus) post-selection in hygromycin, dishes were fixed in methanol, stained with crystal violet, and images were captured.