Hormone-dependent nuclear export of estradiol receptor and DNA synthesis in breast cancer cells -- Data Supplement - JCB--Lombardi et al. -- The Journal of Cell Biology

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Figure S3. The homology between the 444-456 amino acids of ERα and the conserved leucine-rich and REV-like NES of p53. The putative NES sequence of ERα shows homology with the p53 NES (a) and is shared by other steroid receptors (b). (a) The sequence alignment between the indicated regions of ERα and p53 is shown, with the conserved amino acids in red. (b) The sequence alignment between the indicated regions of ERα and other steroid receptors is shown. Sequences were obtained from The National Center for Biotechnology Information Data library: Erα from GenBank/EMBL/DDBJ under accession no. NP000116; Erβ, accession no. NP001035366; PgR, accession no. NP000917; AR, accession no. NP000035; glucocorticoid receptor (GR), accession no. P04150; and mineralocorticoid receptor (MR), accession no. P08235. Conserved amino acids are shown in red; the core of ERα-NES sequence and the putative core of NES sequences in the indicated steroid receptors are underlined. The Tat peptide does not affect EGF-induced p27 nuclear exit in NIH3T3 mouse fibroblasts (c) and does not interfere in subcellular distribution of p53 in MCF-7 cells (d). (c) Quiescent NIH3T3 fibroblasts on coverslips were unstimulated or stimulated for 14 h with 100 ng/ml EGF. When indicated, the Tat peptide (Tat-pep) was added at 1 µM together with EGF. p27 nuclear localization was detected by immunofluorescence as described previously (Castoria, G., A. Migliaccio, M. Di Domenico, M. Lombardi, A. de Falco, L. Varricchio, A. Bilancio, M.V. Barone, and F. Auricchio. 2004. Mol. Cell. Biol. 24:7643–7653) and expressed as the percentage of total cells. (d) Quiescent MCF-7 cells on coverslips were unstimulated or stimulated with 10 nM estradiol for the indicated times. When indicated, the Tat peptide (Tat-pep) was added at 1 µM together with estradiol. Nuclear localization of p53 was revealed by immunofluorescence and expressed as the percentage of total cells. (c and d) Data were derived from at least 1,000 scored cells. The results of several independent experiments were averaged; means and SEM are shown.