Induction of O2− accumulation and apoptosis by 2-ME in CLL cells.
(A) Accumulation of O2− in primary CLL cells treated with 30 μM 2-ME for 24 hours. Primary leukemia cells freshly isolated from patients with CLL were treated with 2-ME in vitro and the O2− levels were determined by flow cytometry analysis as described in “Materials and methods.” (B) Morphologic changes characteristic of apoptosis induced by 2-ME in CLL cells. After cells were incubated with 2-ME under the conditions indicated, the samples were stained and photographed as described in “Materials and methods.” All photographs show microscopic view using × 60 objective. (C) 2-ME induced DNA fragmentation in CLL cells treated with 10 or 30 μM 2-ME for 72 hours. (D) 2-ME induced apoptosis in CLL cells as measured by annexin V reactivity using a flow cytometry analysis. The number shown below each panel indicates the percentage of annexin-positive cells.
