He-GIW induces a necrotic cell death.

<p><b>(a-c)</b> Presence of apoptosis was assayed by FACS analysis of Annexin V/PI dual staining. <b>(a)</b> An example of flow cytometry profile 6 h after treatment with He-GIW or with staurosporine, a positive control for apoptosis. X axis represents annexin-V-APC staining and Y axis PI staining. <b>(b)</b> Percentage of cells positive for Annexin V only (black), PI only (white) or both (grey) was measured 6 h and 24 h after treatment with He-GIW or staurosporine in standard cell culture conditions (c) or with 1% or 5% FCS. <b>(a-c)</b> are representative of 3 independent experiments. <b>(d)</b> Caspase 3 cleavage (red) was assayed by immunofluorescence 6h after He-GIW or staurosporine treatment. Cell nuclei are labeled with Hoechst and appear blue. <b>(e)</b> Caspase 3 cleavage was monitored by Western Blot for 2 h after He-GIW treatment. <b>(f)</b> Adherent cell percentage 6 h after He-GIW treatment in presence or absence of caspase inhibitor Z-VAD. Error bars represent S.E.M. of three independent experiments. p<0.05 (*), p<0.01 (**) and p<0.001 (***)