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. 2014 Sep 12;12:72. doi: 10.1186/s12915-014-0072-6

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© Tin et al.; licensee BioMed Central Ltd. 2014

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Nucleostemin-dependent I3C stimulation of MDM2 nuclear compartmentalization and localization into nucleolus foci. 10AT-Her2 cells were transfected with either control scramble siRNA or nucleostemin siRNA, and then treated with or without 200 μM I3C for 48 hours. (A) The level of nucleostemin protein was determined by Western blot analysis. (B) Cell extracts were biochemically separated into nuclear enriched and cytoplasmic fractions, electrophoretically fractionated, and Western blots probed with antibodies specific for MDM2, the cytoplasmic marker HSP90 and the nuclear marker lamin. (C) The subcellular localization of MDM2 and nucleostemin was determined by indirect immunofluorescence microscopy. DAPI staining was used to visualize DNA stained nuclei. Scale bar represents 4 μm. DAPI, 4',6-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; I3C, indole-3-carbinol; MDM2, murine double mutant 2; NS, nucleostemin; siRNA, small interfering RNA.