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. 2004 Jan 15;23(2):406–417. doi: 10.1038/sj.emboj.7600052

Figure 4.

Figure 4

CRT −/− cells have a short proliferative lifespan. (A) Lifespan of CRT −/− MEF. Primary MEF with indicated genotypes were propagated in culture as described in Materials and methods. Accumulated numbers of population doublings are shown. The data represent experiments using two independent MEF preparations for each genotype. (B) Expression of cyclins A and D1 in WT and CRT-deficient cells during cell cycle progression. Cells were synchronized by serum starvation, stimulated by 10% FBS and harvested at the times shown on the top. In all, 50 μg of proteins were analyzed by SDS–PAGE, followed by immunoblotting with antibodies to CRT, cyclin D1, cyclin A, p21 and β-actin as a loading control. Bar graphs show levels of p21 as a summary of three independent experiments. (C) Reloading of CRT-deficient MEF with CRT increases cell proliferation. Incorporation of BrdU (red) was examined in WT or CRT −/− MEF after nine PDs transfected with His-CRT. DAPI staining shows all cells in the field, and GFP indicates transfected cells. Bar graphs show a summary of three independent experiments.