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PDF file - 174KB, siRNA-mediated knockdown of Noxa and Puma results in a decrease in corresponding transcript levels. CLL cells (N=4) were transfected with two individual siRNA's against Noxa, a Puma-targeting siRNA or control siRNA using Amaxa program X-05. Immediately after nucleofection, cells were cultured on CD40L-expressing stroma for 24 hours, followed by incubation with MLN4924 or vehicle control. (A) Apoptosis within CD19+ subset of cells was determined by Annexin V and 7-AAD staining after 48 hours with MLN4924. (B) Total RNA was isolated from CLL B-cells after 24 hours incubation with MLN4924, reverse-transcribed and subjected to real-time PCR with the indicated probes (in duplicates). Results were normalized to 18S levels. Fold induction of the corresponding mRNA compared to control siRNA-transfected samples is shown. Data are the mean � SE. * - p |