HeLa cells cultured on glass substrata produce numerous prominent focal contacts, which reside at the termini of actin microfilament bundles. However, very few of the cells stain for fibronectin with specific anti-fibronectin antibody. Moreover, the cells form focal contacts in fibronectin-depleted medium, in the presence of high concentrations of anti-fibronectin immunoglobulin G and in the presence of monensin. No fibronectin synthesis can be detected by [35S]methionine-labelling and immunoprecipitation. The possibility that HeLa cell focal contacts are independent of fibronectin in their formation is discussed in relation to the controversy about the relationship between fibronectin and focal contacts.

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