Figure 6

Short-term knockdown of IAPs decreases long-term survival. Cells were transfected with the combination of all three siRNAs targeting cIAP-1 (0.5 nM), cIAP-2 (0.5 nM), and XIAP (0.5 nM) (siTarget), or corresponding non-targeting control siRNA (0.5 nM of each control siRNA to a final concentration of 1.5 nM) (siCon). A, B. 24 hour post-transfection cells were re-seeded for clonogenic assay as described in methods. Cells were grown until colonies formed (2 weeks), then stained, fixed and counted to determine surviving fraction. A. Representative image of colony formation in Control cells (non-targeting siRNA; siCon) and following combined IAP knockdown (siTarget). B. Clonogenic survival in IAP targeted cells. Surviving fraction was calculated as described in methods and expressed as % survival. *p = 0.024 verses siRNA control (siCon), n = 4 individual experiments. C. Expression of proliferation marker ID-1 following IAP knockdown as shown by Western blotting.