CD2-associated protein (CD2AP), an adaptor protein, plays several important roles in podocyte function, linking slit diaphragms to actin-based cytoskeleton and sending survival signals. Here, we investigated whether ginseng total saponin (GTS) had a protective role in the changes of podocyte CD2AP protein and podocyte apoptosis under in vitro diabetic conditions. Conditionally immortalized mouse podocytes cultured with normal glucose (5 mM) or high glucose (30 mM) and with or without advanced glycosylation end products were treated with GTS. We found that CD2AP co-localized with the F-actin fibers in podocyte cytoplasm using confocal imaging; however, diabetic conditions caused the podocytes to diminish and conglomerate CD2AP stainings in the peripheral cytoplasm, which were recovered by GTS. Diabetic conditions also suppressed CD2AP protein levels at 6 and 24 h in western blotting. These phenotypical changes of CD2AP protein were mitigated by GTS. Diabetic conditions also induced podocyte apoptosis at 24 h, which were attenuated by GTS. These findings provide a novel mechanism that diabetic conditions induce quantitative and qualitative changes of podocyte CD2AP protein and apoptosis, which would be restored by GTS.