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Figure 5


Fig. 5. Cell-cell- and cell-substratum-dependent expression of cyclin D1 mRNA is mediated by Rac. MCF10A cells were infected with adenoviruses encoding GFP, β-galactosidase (LacZ), N17Rac or β2-chimerin and serum starved. Control viruses were used at the highest MOI of the test virus. The infected, serum-starved cells were plated on collagen-coated dishes and stimulated with 10% FBS and growth factor cocktail. (A) Cells were mitogen stimulated for 9 hours, and the effect of N17-Rac on cyclin D1 mRNA was determined by QPCR. (B) Cells were mitogen stimulated for 9 hours, and the effect of β2-chimerin on cyclin D1 mRNA was determined by QPCR. (C) MCF10A cells were transfected with control or E-cadherin siRNA and then infected with β-galactosidase (LacZ) or N17-Rac adenovirus during serum- and growth factor-starvation. Quiescent, transfected/infected cells were trypsinized, reseeded in monolayer (Mono) or suspension (Susp) on collagen- or agarose-coated dishes, respectively. The cells were stimulated with 10% FBS and growth factor cocktail for 12 hours. Total RNA was isolated and analyzed for cyclin D1 mRNA by QPCR.





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