Fig. 4.
Aurora A knockdown sensitized CML cells to imatinib treatment. (A) Three days after mock or ShAuroraA infection, 5 × 105 KCL-22 or K562 cells per well were treated with 2.5 or 0.5 μM imatinib (STI), respectively, in 1 ml media in 24-well plate in triplicate and viable cell numbers were counted at days indicated after the drug treatment. (B) Apoptosis analysis of K562 cells. The cells were transduced by mock or ShAuroraA and treated with 0.5 μM imatinib as in (A). Apoptosis of K562 cells was analyzed by annexin V staining after 48 h drug treatment. (C) Relapse assay in liquid culture. Three days after mock or ShAuroraA infection, 5 × 105 KCL-22 cells per well were treated with 2.5 or 5 μM imatinib in 1 ml media in 24-well plate in triplicate and viable cell numbers were counted at indicated days for the prolonged culture. (D) Clonogenic assay for BCR-ABL mutation. Three days after mock or ShAuroraA infection, 1 × 106 KCL-22 cells per well were cultured in standard two-layer soft agar with 5 μM imatinib in triplicate in six-well plate. The numbers of imatinib resistant colonies were scored after 21 days.