The treatment of U-937 human promonocytic cells with the differentiation inducer sodium butyrate (0.75 mM) transiently increased heat-shock protein 70 (HSP70) mRNA levels between 3 and 6 h, and heat-shock protein 27 (HSP27) mRNA levels between 12 and 24 h, as indicated by northern blot assays. Gel retardation assays indicated that butyrate also stimulated heat-shock factor (HSF) binding activity between 3 and 6 h, suggesting that the activation of HSP70 gene expression was mediated by the heat-shock factor DNA response element (HSE). In addition, the treatment provoked a biphasic alteration of the c-fos mRNA level, consisting of a slight increase between 0.5 and 3 h followed by a greater increase between 12 and 48 h, while it caused a single increase between 12 and 48 h in c-jun mRNA level. The possible involvement of the heat-shock protein genes in the butyrate-induced differentiation of U-937 cells is discussed.