Objective To study the distribution of CD2-associated protein (CD2AP) in normal renal cell lines and its interaction with nephrin and F-actin in podocytes. Methods The human mesangial cells (HMC) and HK-2 were cultured in DMEM. Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 The expressions of CD2AP and nephrin in podocytes were examined by RT-PCR and Western blotting. The distribution of CD2AP in HMC, HK-2, differentiated and undifferentiated podocytes was observed by laser scanning confocal microscopy. The coexistence of CD2AP with nephrin and F-actin in undifferentiated podocytes were also detected. Results CD2AP was distributed within the cytoplasm and perinulcear region of HK-2 and undifferentiated podocytes, but was absent in HMC cells. Its distribution profile changed and presented as peripheral accumulation when podocytes were put into differentiationpermissive conditions. CD2AP was located together with nephrin and F-actin in podocytes. Conclusion CD2AP can be detected in epithelial-originated renal cells. The alteration of distribution profile of CD2AP indicates it may participate in the process of podocytes differentiation and be involved in the regulation of slit diaphragm and cytoskeleton.