Fig. 8. dRheb is required in cultured cells for G1/S progression and cell growth. The effect of inhibiting dRheb expression by dsRNA was tested in S2 cells transfected with pPAC-FLAG or pPAC-FLAG-dRheb. After 72 hours, dsRNA-treated or untreated cells were lysed. Western-blot analysis of total protein with anti-FLAG antibody revealed inhibition of FLAG-dRheb expression in the presence of dRheb dsRNA (A). When cells treated with dRheb dsRNA were harvested one to eight days after treatment and subjected to flow cytometry, they were found to gradually arrest at G0/G1 (B). Forward scatter analysis of cells collected two, three, four and five days after the addition of dRheb dsRNA shows that it causes a reduction in cell size (C).

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