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Fig. 4. Knockdown of both topo II isoforms by siRNAs. (A) HeLa cells were transfected with both {alpha} siRNA-1 and ß siRNA-2 (1; 5 µl each siRNA was added per 35 mm dish; 2; 10 µl each) or buffer as a control. Cells were assayed on the third or fourth day after transfection by immunoblotting using the anti-topo II{alpha} antibody 8D2 (left) and the anti-topo IIß antibody 3B6 (right). (B) The topo II{alpha}ß-knockdown cells were analyzed using the LSC2 3 days after transfection. (C) Topo II{alpha}ß-knockdown cells (3 days after transfection) were stained with DAPI, anti-topo II{alpha}ß antibody (6H8) and anti-tubulin antibody (YL1/2). Bar in C, 20 µm (upper panels) and 10 µm (lower panels).