140 | Chapter 5 Supplemental Figure 2. RAG2 mRNA expression in BV173 human pre-B ALL cell line and WT mouse Abl pre-B cells. Rag2 real-time RT-PCR analysis of the human BCR-ABL+ B-ALL cell line BV173 and WT mouse Abl pre-B cells. Cells were treated with 2.5 mM IKKβi, 2.5 mM AKTi or 10 mM STI571 for 48h. Means ± SD are shown (n=4 independent stimulations, PCR reactions were performed in duplicate), *p<0.05, as determined by the one sample t test. Supplemental Figure 3. Combined treatment with IKKβi and AKTi inhibits NF-kB transcription factor activity in human pre-B ALL cell lines. BV173 and SUP-B15 cells were treated for 24h with 2.5 mM IKKβi, 2.5 mM AKTi, or with both. Equal amounts of nuclear lysates (5 mg) were analyzed for specific NF-kB p65 (white bars) and p50 (grey bars) transcription factor DNA binding activity using an enzyme-linked immunosorbent assay (ELISA) (Abcam ab133128). Each stimulation was performed in triplicate and each individual lysate was assayed in duplicate. Means ± SD of independent cultures are shown (n=3). *p<0.05, as determined by the one sample t test.
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