Serotonin 5-HT(2B) receptors are often coexpressed with 5-HT(1B) receptors, and cross-talk between the two receptors has been reported in various cell types. However, many mechanistic details underlying 5-HT(1B) and 5-HT(2B) receptor cross-talk have not been elucidated. We hypothesized that 5-HT(2B) and 5-HT(1B) receptors each affect the others' signaling by modulating the others' trafficking. We thus examined the agonist stimulated internalization kinetics of fluorescent protein-tagged 5-HT(2B) and 5-HT(1B) receptors when expressed alone and upon coexpression in LMTK(-) murine fibroblasts. Time-lapse confocal microscopy and whole-cell radioligand binding analyses revealed that, when expressed alone, 5-HT(2B) and 5-HT(1B) receptors displayed distinct half-lives. Upon coexpression, serotonin-induced internalization of 5-HT(2B) receptors was accelerated 5-fold and was insensitive to a 5-HT(2B) receptor antagonist. In this context, 5-HT(2B) receptors did internalize in response to a 5-HT(1B) receptor agonist. In contrast, co-expression did not render 5-HT(1B) receptor internalization sensitive to a 5-HT(2B) receptor agonist. The altered internalization kinetics of both receptors upon coexpression was probably not due to direct interaction because only low levels of colocalization were observed. Antibody knockdown experiments revealed that internalization of 5-HT(1B) receptors (expressed alone) was entirely clathrin-independent and Caveolin1-dependent, whereas that of 5-HT(2B) receptors (expressed alone) was Caveolin1-independent and clathrin-dependent. Upon coexpression, serotonin-induced 5-HT(2B) receptor internalization became partially Caveolin1-dependent, and serotonin-induced 5-HT(1B) receptor internalization became entirely Caveolin1-independent in a protein kinase Cepsilon-dependent fashion. In conclusion, these data demonstrate that coexpression of 5-HT(1B) and 5-HT(2B) receptors influences the internalization pathways and kinetics of both receptors.