Phlorizin, 0.5 mM, increases the uptake of tritiated p-aminohippuric acid (PAH) in rat kidney cortex slices in vitro. Phlorizin also diminishes the rate of 3H-PAH washout from preloaded slices into PAH-free medium. At higher concentrations, phlorizin (5.0 mM) reduces slice uptake of 3H-PAH following short incubations but increases 3H-PAH accumulation after more prolonged incubations. Section freeze-dry autoradiography demonstrates that phlorizin inhibits secretion of 3H-PAH from cell to lumen in proximal tubules. Consequently, the increased 3H-PAH uptake and delayed washout induced by phlorizin may be attributed to effects at the antiluminal cell membrane. Phlorizin stimulation of PAH uptake occurs despite inhibition of secretion across the luminal membrane. Intracellular accumulation of 3H-phlorizin, demonstrable by autoradiography, provides direct evidence that cellular accumulation affords the glycoside access to both the luminal and antiluminal membrane in proximal tubules. These interactions between phlorizin and PAH suggest shared features of the membrane transport systems for secretion and reabsorption of sugars and organic acids in kidney.