Laminin promotes attachment and process formation in the neuroblastoma X glioma hybrid cell line NG108-15. As cells attached to laminin, they flattened and remained dispersed rather than associated in clumps. Process formation was observed within 1 hr after exposure to laminin and was dose dependent. Cycloheximide, an inhibitor of protein synthesis, did not block laminin-mediated attachment and neurite formation. Addition of drugs that depolymerize the cytoskeleton led to different behaviors for cells grown on plastic compared with those in the presence of laminin. Cells on plastic treated with either vinblastine or cytochalasin neither flattened nor grew processes. Cells plus laminin and vinblastine retracted processes, but remained flat, suggesting that laminin-induced processes can be destabilized by disrupting microtubules. Cells sequentially treated with laminin and cytochalasin produced processes that were thin and highly branched. Cells in high concentrations of cytochalasin on a laminin substrate formed aberrant processes even when their soma did not flatten. Since laminin counteracted the effect of cytochalasin on process outgrowth but did not alter the effect of cytochalasin on flattening of the cell body, different mechanisms mediated by microfilaments may be involved in cell flattening and in process formation.