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. Author manuscript; available in PMC: 2011 Feb 2.
Published in final edited form as: Cell Cycle. 2009 Jan 1;8(1):125–136. doi: 10.4161/cc.8.1.7528

Figure 1.

Figure 1

Acute depletion of Mcm7 inhibits S-phase entry in primary HDF. Quiescent hTERT-expressing HDF were transfected with siMcm7 or siCon oligonucleotides, then stimulated to enter the cell cycle. 24 hours after serum-stimulation, chromatin and soluble fractions were prepared and analyzed by SDS-PAGE and immunoblotting for expression of various proteins as indicated (A). Parallel cultures of siCon and siMcm7-transfected cells were incubated with 25 μM BrdU for 1 hr. The cell cycle distributions of the resulting HDF were analyzed by flow cytometry, as described under ‘Materials and Methods’ (B). In a similar experiment, quiescent HDF were transfected with siMcm7 or siCon oligonucleotides, then stimulated to enter the cell cycle. After 0, 12 and 15 hours of serum stimulation, cells were lysed and CDK4 and CDK2 complexes were immunoprecipitated for in vitro kinase assays. Rb-GST and histone H1 were used as substrates for CDK4 and CDK2 respectively (C).