A major aim of this investigation was to determine whether, in steady-state ascites cells, Cl- transport can be partitioned into a furosemide-sensitive cotransport with K+ and a separate 4,4'-isothiocyanostilbene-2,2'-disulfonic acid (DIDS) sensitive self-exchange. Both Cl- and K+ fluxes were studied. The furosemide- and Cl- sensitive K+ fluxes were equivalent, both in normal ionic media and when the external K+ concentration, [K+]o, was varied from 4 to 30 mM. The stoichiometry of the furosemide-sensitive Cl- and K+ fluxes was 2 Cl-:1 K+ at 0.1 and 0.5 mM drug levels but increased to 3 Cl-:1 K+ at 1.0 mM furosemide. DIDS at 0.1 mM had no effect on the K+ exchange rate but inhibited Cl- exchange by 39% +/- 2 (S.E.). The effects of DIDS and 0.5 mM furosemide on Cl- transport were additive but 1.0 mM furosemide and DIDS had overlapping inhibitory actions. Thus furosemide acts on components of K+ and Cl- transport which are linked to each other, but the drug also inhibits an additional DIDS-sensitive Cl- pathway, when present at higher concentrations. The dependence of the furosemide-sensitive K+ and Cl- transport on [K+]o was also studied; both fluxes fell as the [K+]o increased. The latter results recall those in an earlier study by Hempling (Hempling, H.G. (1962) J. Cell. Comp. Physiol. 60, 181-198).