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By E.Y. C. Lee

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MILAM 10 min. All the cells were detached. After collection by centrifugation each pellet was suspended in 2 ml adhesion medium (5% FCS) and turbidity measured at 6^+0 nm. Fig. e. cells with no additions which show a uniform field of cells having only few small clumps of cells. 30 BIOLOGY AND CHEMISTRY OF EUCARYOTIC CELL SURFACES Fig· lb shows cells grown in the presence of 1 mM theophylline for 2k hrs. In this case there are large aggregates of cells with only a few small aggregates. The agglutinability of cells grown in the presence of theophylline, glucagon or insulin was shown to he time dependent and also dependent on the concentration of the material added.

Fab absorbed with such mutants should still be able to fully inactivate the contact-site-A system, because blockage of either a_ or <*c_ would be sufficient. 3). This conclusion is independent of the number of sub-classes in the system. However, if the same Fab preparation is absorbed with two mutants which have no defect in common, the double absorption would remove the full contact-site-A blocking activity, as absorption with wild-type cells does. The Fab-absorbing activity of non-aggregating mutants has been tested in order to demonstrate heterogeneous components of the contact-site-A system (28).

Other metabolic poisons (F~, N3" and cycloheximide) did not inhibit adhesion. We have also found that attached cells bind less N-ethylmaleimide than free cells and the inhibition of adhesiveness was reduced. A divalent metal requirement for maintenance of intercellular adhesion originally was shown by the dissociation of frog embryos in Ca 2+ free medium. The disruption of cell aggregates by EDTA is attributed to the metal requirement for formation and stabilization of cell aggregates although other factors may also be involved (Gingell et dl.

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