Gelsolin and caspase-3 were cleaved during apoptosis induced by Fas ligation.
(A) Transfected Jurkat cells were incubated for the indicated period of time with 100 ng/mL CH11 anti-Fas mAb. Immunoblot analysis of whole-cell lysates (1 × 106 cell equivalents) demonstrated overexpression of gelsolin and its cleavage during apoptosis. The gelsolin cleavage product is evident at approximately 46 kd. Results are representative of 4 independent experiments. (B) Parallel samples of transfected Jurkat cells incubated with 100 ng/mL CH11 anti-Fas mAb were assayed for the percentage of cell deaths by trypan blue exclusion (closed circles, cells overexpressing gelsolin, JK-Gsn-22; open circles, vector controls, JK-Vec-33). The percentage of cell deaths of parallel samples was confirmed by staining with Annexin V-FITC (solid bars, JK-Gsn-22; hatched bars, JK-Vec-33). (C) Transfected Jurkat cells were treated with anti-Fas mAb 100 ng/mL CH11 for the indicated periods of time. Immunoblot analysis of postnuclear lysates (1 × 106 cell equivalents) demonstrated cleavage of caspase-3; one of the fragments of caspase-3 generated during apoptosis is evident at approximately 17 kd. Results are representative of 3 independent experiments.
