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Blood, Vol. 112, Issue 9, 3827-3834, November 1, 2008
p53-mediated apoptosis of CLL cells: evidence for a transcription-independent mechanism
Blood Steele et al.
112: 3827
Supplemental materials for: Steele et al
Files in this Data Supplement:
- Figure S1. DDF analysis of distribution of p53 and marker proteins in CLL cells from patient 6 following treatment for 18h with 10 µM nutlin 3a in the presence of 100 µM ZVAD (JPG, 62.4 KB)
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Fraction 3 samples were analyzed on a separate western blot which was exposed to the same antibody solutions as were Fraction 1 and 2 samples. Lanes have been juxtaposed for clarity of presentation.
- Figure S2. Downregulation of chlorambucil-induced p53 targets by PFTα (JPG, 36.5 KB)
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Cells from patient 19 were incubated in the presence of 100 µM ZVAD with chl and/or PFTα as indicated. Cells were extracted and analysed by western blotting.
- Figure S3. Cells were incubated with fludarabine, nutlin, or PFTα as indicated, extracted, and analysed by western blotting (JPG, 67.1 KB)
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Cells from patient 18 were analysed in the same experiment to provide a positive control for the immunodetection of p53, MDM2, and p21CIP1.
- Figure S4. Augmentation of fludarabine-induced apoptosis by PFTα (JPG, 48.3 KB)
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The percentage of apoptotic cells, clearly identified by nuclear condensation, were computed and are displayed as a histogram in Fig. 4B.
- Figure S5. PFTα augments apoptosis induction by clinically achievable concentrations of cytotoxic drugs (JPG, 52.3 KB)
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Cells from patient 28 were incubated with cytotoxic drugs and/or PFTα as indicated. Apoptosis was quantified by western blot analysis of PARP cleavage (A) and by morphological assessment (B). The data are typical of seven independent experiments, carried out using cells from patients 22 and 25–30.
- Figure S6. Cells from patient 26 were incubated with chl, nutlin 3a, and/or PFTα exactly as described in the legend to Fig. 6, except that ZVAD was omitted (JPG, 34.4 KB)
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Apoptiosis induction was quantified by western blot analysis of PARP cleavage (A) or by morphology (B).
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