ABSTRACT
Cell adhesion and mitogenic growth factors are jointly required for cell proliferation of almost all cell types. Cell proliferation reflects continued progression through the cell cycle, and progression through the cell cycle is mediated by the sequential activation of cyclin-dependent kinases (cdks). Cell cycle progression through G1 phase is mediated by a subset of the cyclin-dependent kinases: cyclin D-cdk4 or cyclin D-cdk6, cyclin E-cdk2, and cyclin A-cdk2. This chapter describes the methods for analyzing the regulation of G1 phase cyclins, cdk inhibitors, and cdk activities. It stimulates cell cycle progression in monolayer, the trypsinized cells are seeded in untreated tissue culture dishes in serum-containing medium or in matrix-protein coated dishes. Trypsin-dependent activation of ERKs should be considered when setting up experiments designed to examine time- or anchorage-dependent ERK activation.
