sequential ATP-dependent events involving first hsp70 and then hsp90 in opening of the steroid binding pocket. J Biol Chem 275: 18054–18060. 53. Murphy PJ, Kanelakis KC, Galigniana MD, Morishima Y, Pratt WB (2001) Stoichiometry, abundance, and functional significance of the hsp90/hsp70- based multiprotein chaperone machinery in reticulocyte lysate. J Biol Chem 276: 30092–30098. 54. Dittmar KD, Demady DR, Stancato LF, Krishna P, Pratt WB (1997) Folding of the glucocorticoid receptor by the heat shock protein (hsp) 90-based chaperone machinery. The role of p23 is to stabilize receptor.hsp90 heterocomplexes formed by hsp90.p60.hsp70. J Biol Chem 272: 21213–21220. 55. Dauvois S, White R, Parker MG (1993) The antiestrogen ICI 182780 disrupts estrogen receptor nucleocytoplasmic shuttling. J Cell Sci 106( Pt 4): 1377–1388. 56. Savory JG, Hsu B, Laquian IR, Giffin W, Reich T, et al. (1999) Discrimination between NL1- and NL2-mediated nuclear localization of the glucocorticoid receptor. Mol Cell Biol 19: 1025–1037. 57. Hache RJ, Tse R, Reich T, Savory JG, Lefebvre YA (1999) Nucleocytoplasmic trafficking of steroid-free glucocorticoid receptor. J Biol Chem 274: 1432–1439. 58. Kang KI, Devin J, Cadepond F, Jibard N, Guiochon-Mantel A, et al. (1994) In vivo functional protein-protein interaction: nuclear targeted hsp90 shifts cytoplasmic steroid receptor mutants into the nucleus. Proc Natl Acad Sci U S A 91: 340–344. 59. Martins VR, Pratt WB, Terracio L, Hirst MA, Ringold GM, et al. (1991) Demonstration by confocal microscopy that unliganded overexpressed gluco- corticoid receptors are distributed in a nonrandom manner throughout all planes of the nucleus. Mol Endocrinol 5: 217–225. 60. Sanchez ER, Hirst M, Scherrer LC, Tang HY, Welsh MJ, et al. (1990) Hormone-free mouse glucocorticoid receptors overexpressed in Chinese hamster ovary cells are localized to the nucleus and are associated with both hsp70 and hsp90. J Biol Chem 265: 20123–20130. 61. Daniel C, Albrecht H, Lu¨ dke A, Hugo C (2008) Nestin expression in repopulating mesangial cells promotes their proliferation. Lab Invest Feb 11 [Epub ahead of print]. 62. Son GH, Geum D, Jung H, Kim K (2001) Glucocorticoid inhibits growth factor- induced differentiation of hippocampal progenitor HiB5 cells. J Neurochem 79: 1013–1021. 63. Sousa N, Paula-Barbosa MM, Almeida OF (1999) Ligand and subfield specificity of corticoid-induced neuronal loss in the rat hippocampal formation. Neuroscience 89: 1079–1087. 64. Miller AL, Komak S, Webb MS, Leiter EH, Thompson EB (2007) Gene expression profiling of leukemic cells and primary thymocytes predicts a signature for apoptotic sensitivity to glucocorticoids. Cancer Cell Int 7: 18. Nestin and Steroid Receptor 65. Ramalingam A, Hirai A, Thompson EA (1997) Glucocorticoid inhibition of fibroblast proliferation and regulation of the cyclin kinase inhibitor p21Cip1. Mol Endocrinol 11: 577–586. 66. Buse P, Woo PL, Alexander DB, Cha HH, Reza A, et al. (1995) Transforming growth factor-alpha abrogates glucocorticoid-stimulated tight junction formation and growth suppression in rat mammary epithelial tumor cells. J Biol Chem 270: 6505–6514. 67. Vanderbilt JN, Miesfeld R, Maler BA, Yamamoto KR (1987) Intracellular receptor concentration limits glucocorticoid-dependent enhancer activity. Mol Endocrinol 1: 68–74. 68. Cintra A, Solfrini V, Bunnemann B, Okret S, Bortolotti F, et al. (1993) Prenatal development of glucocorticoid receptor gene expression and immunoreactivity in the rat brain and pituitary gland: a combined in situ hybridization and immunocytochemical analysis. Neuroendocrinology 57: 1133–1147. 69. Matthews SG (1998) Dynamic changes in glucocorticoid and mineralocorticoid receptor mRNA in the developing guinea pig brain. Brain Res Dev Brain Res 107: 123–132. 70. Bird AD, Tan KH, Olsson PF, Zieba M, Flecknoe SJ, et al. (2007) Identification of glucocorticoid-regulated genes that control cell proliferation during murine respiratory development. J Physiol 585: 187–201. 71. Wintermantel TM, Bock D, Fleig V, Greiner EF, Schutz G (2005) The epithelial glucocorticoid receptor is required for the normal timing of cell proliferation during mammary lobuloalveolar development but is dispensable for milk production. Mol Endocrinol 19: 340–349. 72. Reagan LP, McEwen BS (1997) Controversies surrounding glucocorticoid- mediated cell death in the hippocampus. J Chem Neuroanat 13: 149–167. 73. Peschle C, Botta R, Mu¨ller R, Valtieri M, Ziegler BL (2001) Purification and functional assay of pluripotent hematopoietic stem cells. Rev Clin Exp Hematol 5: 3–14. 74. Bonanno G, Perillo A, Rutella S, De Ritis DG, Mariotti A, et al. (2004) Clinical isolation and functional characterization of cord blood CD133+ hematopoietic progenitor cells. Transfusion 44: 1087–1097. 75. Yaworsky PJ, Kappen C (1999) Heterogeneity of neural progenitor cells revealed by enhancers in the nestin gene. Dev Biol 205: 309–321. 76. Zimmerman L, Parr B, Lendahl U, Cunningham M, McKay R, et al. (1994) Independent regulatory elements in the nestin gene direct transgene expression to neural stem cells or muscle precursors. Neuron 12: 11–24. 77. Neidhardt J, Fehr S, Kutsche M, Lohler J, Schachner M (2003) Tenascin-N: characterization of a novel member of the tenascin family that mediates neurite repulsion from hippocampal explants. Mol Cell Neurosci 23: 193–209. 78. Yang J, Cheng L, Yan Y, Bian W, Tomooka Y, et al. (2001) Mouse nestin cDNA cloning and protein expression in the cytoskeleton of transfected cells. Biochim Biophys Acta 1520: 251–254. PLoS ONE | www.plosone.org 16 June 2009 | Volume 4 | Issue 6 | e6084