Figure 5. CD59 inhibition sensitizes cancer cells that undergo EMT to cetuximab mediated CDC.
A549 and H358 cell lines transfected either with control or CD59 siRNA (50 nM) were serum starved for 24 h and stimulated with or without TGF-β (5 ng/ml) for 72 h. To assess complement dependent cytotoxicity, cells were incubated with normal human serum (NHS) or heat inactivated serum (HI) in the presence or absence of cetuximab, fixed in 70% alcohol before staining with 7-AAD. Cituximab alone or IgG are used a controls. DNA fragmentation associated with complement dependent cell lysis was assessed by the percentage of 7-AAD positive cells in sub-G0 by flow cytometry (A & B).