The pp60c-src is present at high level in differentiated retinal pigment epithelium (RPE) in culture. Immunofluorescence microscopy using GD11 (anti-pp60c-src) shows intense staining in the plasma membrane, especially at the cell-cell junctions, and diffuse staining in the cytoplasma. Western blot analysis shows that the majority of the GD11-reactive molecules is localized in the membrane. The pp60c-src does not translocate between membrane and cytoplasma when the RPE was reacted with vasoactive intestinal peptide (VIP), which is previously shown to stimulate phosphorylation of the pp60c-src in the membranes (Biochem. Biophys. Res. Commun. 174, 452-8, 1991). Here, VIP modulation is shown to alter the reactivity of pp60c-src with a monoclonal anti-phosphotyrosine antibody. © 1992 Academic Press Ltd.