AD 198 induced apoptosis in TRAF3
tumor B cells. (A and B) Cell cycle distribution determined by PI staining and flow cytometry. TRAF3-/- mouse B lymphoma cell lines (A) or human MM cell lines (B) were cultured in the absence or presence of AD 198 of indicated concentration for 24 h, and then fixed. Fixed cells were stained with PI and analyzed by FACS. Representative histograms of PI staining are shown, and percentage of apoptotic cells (DNA content < 2n; sub-G1) and proliferating cells (2n < DNA content ≤ 4n; S/G2/M) are indicated. Results are representative of 3 independent experiments. (C) Western blot analysis of caspase 3 cleavage. Cells were cultured in the absence or presence of AD 198 of indicated concentration. Total cellular lysates were prepared at indicated time points, and then immunoblotted for caspase 3, followed by actin. Immunoblots of actin were used as loading control. Results are representative of three independent experiments. Similar results were also obtained with other cell lines examined.