Figure 4. Lunasin's anti-proliferative activity on NSCLC cells is not apoptosis-induced.

Apoptosis determination in H661 cells was ascertained microscopically by the detection of externalized phosphatidylserine bound by a fluorescent annexin conjugate under anchorage-dependent growth conditions. Cells were grown in the presence of vehicle or 100 μM lunasin for 24 hours or 1 μM staurosporine for 4 to 6 hours (positive control). A) Representative 10X bright-field images showing the absence of characteristic apoptotic morphology in cells treated for 24 hours with lunasin in contrast to the typical apoptotic morphological features, rounding and blebbing, exhibited by cell exposed to staurosporine for 6 hours. Scale bar represents 50 μm. B) Representative fluorescent 40X images showing the absence of annexin detection and lack of characteristic apoptotic morphology in cells exposed to lunasin for 24 hours in contrast to positive annexin detection and the onset of apoptotic morphology (rounding) in cells exposed to staurosporine for 4 hours. Images were analyzed with AxioVision software v4.6.3.0.