Fludarabine nucleoside induces accumulations of p53, p63 and p73 in the nuclei of human B-lymphoid cell lines, with cytosolic and mitochondrial increases in p53
Article first published online: 29 JUN 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PROTEOMICS - Clinical Applications
Volume 6, Issue 5-6, pages 279–290, June 2012
How to Cite
Almazi, J. G., Mactier, S., Best, O. G., Crossett, B., Mulligan, S. P. and Christopherson, R. I. (2012), Fludarabine nucleoside induces accumulations of p53, p63 and p73 in the nuclei of human B-lymphoid cell lines, with cytosolic and mitochondrial increases in p53. Prot. Clin. Appl., 6: 279–290. doi: 10.1002/prca.201200003
- Issue published online: 29 JUN 2012
- Article first published online: 29 JUN 2012
- Accepted manuscript online: 29 MAY 2012 02:04AM EST
- Manuscript Accepted: 6 APR 2012
- Manuscript Revised: 16 MAR 2012
- Manuscript Received: 12 JAN 2012
Disclaimer: Supplementary materials have been peer-reviewed but not copyedited.
Figure S1: 2D Western blots for phospho-serine 15 and phospho-serine 392 of p53 in whole lysates of Raji cells treated with 2-FaraA (3 μM) for control, 24 h and 48 h treated samples. Serine 15 is phosphorylated in control and 2-FaraA treated cells, and remains phosphorylated in the low molecular weight derivatives. Full length p53 is phosphorylated at Serine 392 after treatment with 2-FaraA, but not in control cultures. However, treatment of 2-FaraA does not induce phosphorylation of low molecular weight derivatives at Serine 392. Extracts (100 μg protein) were focused on an 11 cm pH 3-10 IPG strip and separated in the second dimension using 4-12% SDS-PAGE. Proteins were then transferred onto a PVDF membrane and Western blotting was performed with the p53 antibodies (#9284S) and (#9281S).
Figure S2: 2D Western blots for p53 in whole lysates of Raji cells treated with 2-FaraA (3 μM ) ± the pan caspase inhibitor Z-VAD-FMK (100 μM) for control, 24 h and 48 h treated samples. Western blotting is described in Materials and Methods. A p53 derivative of 25 kDa is indicated by an arrow for 2-FaraA-treated cells without Z-VAD-FMK.
Figure S3: 1D Western blots for p53 family members in whole lysates of different cell lines treated with 2-FaraA (3 μM). Cells were harvested and lysed in Laemmli buffer. Changes in protein levels can be compared with changes in mRNA levels in Figure 8.
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