We examined the effects of the phorbol ester phorbol 12-myristate 13-acetate (PMA) on the rate of hexose transport into 3T3-L1 adipocytes. Exposure of adipocytes to PMA (1 microM) for 60 min results in a 1.7-2.5-fold increase in the rate of hexose transport. This effect was mediated by translocation of two isoforms of glucose transporters to the plasma membrane, as determined by labelling in situ, photoaffinity labelling with a membrane-impermeant glucose analogue, and by immunoblotting of subcellular fractions. The PMA-induced stimulation of both transport and transporter translocation was substantially less than that induced by insulin in this cell line; the PMA-induced increase in plasma-membrane GLUT 1 and GLUT 4 transporter isoforms was only about 40% and 10% respectively of that induced by insulin. We suggest that the stimulation of transport by insulin and PMA occurs via different mechanisms, which is manifested by the ability of insulin to induce a much greater increase in the plasma-membrane content of GLUT 4 compared with the phorbol ester.