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Figure 7


Fig. 7. Causal relationship between the stimulatory effects of E-cadherin-mediated cell-cell adhesion on cyclin D1 gene expression and S-phase entry. (A) Serum-starved control and E-cadherin (E-cad) siRNA-transfected MCF10A cells were trypsinized, reseeded in full maintenance medium (see Materials and Methods) on collagen-coated, 25-mm glass coverslips at 2x104 cells/cm2 (with cell-cell adhesion) or 2x103 cells/cm2 (without cell-cell adhesion) as described by Liu et al. (Liu et al., 2006). BrdU was added when the cells were seeded and stimulated; the number of BrdU-labeled nuclei (relative to DAPI-stained nuclei) was determined after 24 hours. (B) Duplicate samples were collected at 12 hours and analyzed for cyclin D1 mRNA by QPCR. Results were calculated relative to the level of cyclin D1 mRNA in serum-starved cells expressing control siRNA. (C) Serum-starved MCF10A cells infected with Ad-GFP or Adcyclin D1 (100 MOI) were treated and analyzed as in A. Results for each panel show means ± s.e.m. of two experiments.





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