Effect of AG490 on G-CSF–induced up-regulation of cIAP2 protein and antiapoptosis.
(A) Neutrophils were preincubated with AG490 (50 μM) for 20 minutes, and thereafter cultivated in the presence or absence of G-CSF (50 ng/mL) for 1 hour. The expression of cIAP1 and cIAP2 proteins was analyzed by immunoblotting using antibodies against each protein. As a control, p38 was used. The ratio of cIAP2 to p38 band intensity was calculated, and the ratio at hour 0 is expressed as 1 (lower panel). The cell lysates equivalent to 2.5 × 106 cells were loaded onto each lane. The results shown are representative of 3 independent experiments. (B) Neutrophils were preincubated with AG490 (50 μM) for 20 minutes, and thereafter cultivated in the presence or absence of G-CSF (50 ng/mL). After cultivation for 9 or 24 hours, apoptotic cells were determined by flow cytometry. The data are expressed as means ± standard deviations of 3 experiments. * and ** indicate significantly inhibited by AG490 (*P < .05; **P < .01).
