Secretory cells employ regulated exocytosis to secrete material in the extracellular space. If secretory material is composed of non-soluble components, such as lipids or proteins, it does not readily diffuse from the secretory vesicle after fusion with the plasma membrane and additional forces are required for efficient release. We have recently reported that lamellar bodies (LBs) - secretory vesicles storing pulmonary surfactant in alveolar type II cells are coated with actin upon exocytic fusion with the plasma membrane. Subsequent compression of the actin coat is necessary to extrude lipid-rich surfactant from the vesicle. This compression is facilitated by non-muscle myosin II. Here we investigate the role of other myosins in surfactant secretion, especially the myosin I family. These unconventional myosins contain an actin-binding head domain and a tail which binds to anionic phospholipids such as phosphatidylinositol-4,5-bisphosphate (PIP₂). We overexpressed fluorescently-tagged PIP₂-binding pleckstrin homology (PH) domain of phospholipase C and several members of the myosin I family in primary rat alveolar type II cells and investigated the process of exocytosis using high-resolution, time-lapsed live-cell fluorescence imaging. The fusion of LBs with the plasma membrane was followed by recruitment of the PH domain to the vesicle membrane, indicating the presence of PIP₂ on the vesicle. Several members of the myosin I family also translocate to the vesicle membrane after fusion. Their capability to attach the newly formed actin coat around the vesicle to the PIP₂-enriched vesicle membrane suggests that myosin I plays a role for post-fusion processes in surfactant exocytosis.