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Fig. 3. Disturbed post-mitotic actin reorganization inhibits integrin-mediated focal adhesion assembly, autophosphorylation of FAK and growth factor-stimulated MAPK phosphorylation in early G1 phase. (A) Synchronized N2A cells were incubated with or without CCD and fixed 3 hours thereafter. Vinculin association with actin filaments was determined by fluorescence microscopy. Bars, 20 µm. (B) p42/p44 MAPK phosphorylation and (Y397) FAK autophosphorylation were investigated by western blotting in lysates of mitotic and post-mitotic cells treated for up to 3 hours with CCD or LB (p42 MAPK=loading control). Representative results out of at least three independent experiments are shown. (C) Synchronized cells were incubated for 3 hours with CCD or LB as in (B), after which they were washed once and released for 1 hour in fresh medium. Phosphorylation of FAK and p42/p44 MAPK was then analyzed by western blotting. p42 MAPK, loading control; R, recovery.
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