Supplemental Figure 1:Expression of topo II and topo II protein in HL-60, NB4 or KG1 cells
transfected with si-RNAs targeted to GFP (si-GFP), topo II (si-topo II) or topo II (si-topo II
or si-topo II-L), and in HL-60/AMSA-R cells. Serial dilutions of cell lysates were subjected to
Western blot analysis with a topo II, topo II or -tubulin or actin specific antibody.
Comparison of protein levels of (A) topo II and topo II in HL-60/si-GFP and HL-60/si-topo
II cells; (B) topo II and topo II in HL-60/si-GFP and HL-60/si-topo II cells; (C) topo II
and topo II in HL-60/si-GFP, HL-60/si-topo II-L and HL-60/AMSA-R cells; (D) topo II and
topo II in NB4/si-GFP and NB4/si-topo II cells; and (E) topo II in KG1/si-GFP and KG1/sitopo II cells.
Supplemental Figure 2: Induction of apoptosis in ATRA-treated HL-60/si-topo II cells
involves activation of the caspase cascade. (A) Two aliquots each of HL-60/si-topo II cells
were treated with 0.1% ethanol (-) or 1 M ATRA (+) for 72 h. During the ethanol or ATRA
treatment DMSO was added at 36 h, 48 h and 60 h to one aliquot of the ethanol or ATRA treated
cells, whereas the caspase inhibitor, Q-VD-OPH, was added at 36 h, 48 h and 60 h to the second
aliquot of the ethanol or ATRA treated cells. Apoptosis was determined at 72 h after treatment
with ethanol or ATRA was initiated. Data are the mean of triplicate experiments ± SD. (B) Cell
lysates of HL-60/si-GFP or HL-60/si-topoII cells treated continuously with 0.1% ethanol
(control) or 1 M ATRA for 72 h were subjected to Western blot analysis with an antibody
specific for cleaved caspase 3 (large fragment 17/19kDa of activated caspase 3) or actin specific
antibody.