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The resulting pellets were further homogenized in the Tris salt buffer with the addition of 210% SDS, transferred to ultracentrifuge tubes and spun at 163, 200 g and 10 C for 30 min to extract the insoluble fraction

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Remarkably, ER-, but not ER-specific ligands, lead to increased phosphorylation of ERK 1/2, which initiates cell cycle entry (demonstrated by the up-regulation of mitotic markers such as CDK1/cdc2 and PCNA), DNA replication, and duplication of the centrosome -the major microtubule-organizing center in the cell- in dividing cells (Wang et al., 2008)