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Open Access Research article

Oestrogen blocks the nuclear entry of SOX9 in the developing gonad of a marsupial mammal

Andrew J Pask12*, Natalie E Calatayud2, Geoff Shaw2, William M Wood1 and Marilyn B Renfree2

Author Affiliations

1 Department of Molecular and Cellular Biology, University of Connecticut, Storrs, CT 06260, USA

2 Department of Zoology, The University of Melbourne, Melbourne, Victoria 3010, Australia

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BMC Biology 2010, 8:113  doi:10.1186/1741-7007-8-113

Published: 31 August 2010

Additional files

Additional file 1:

Statistical difference between male and female data points shown in Figure 1. P values (from t-tests) are listed for all data points shown in the normal expression profiles for SOX9, AMH, FGF9, FOXL2, FST, RSPO1, WNT4. Gene name is given in the top left hand corner of each data set. Column 2 shows the stage of development; d = day of gestation, D = postnatal day. Column 3 shows the P value from a two tailed, homoscedastic t-test (conducted in Microsoft Excel). Significant values (P ≤ 0.05) are highlighted in yellow. The bottom row (combined M (male) v F (female)) shows the P values for the entire male versus female data set combined across the time period shown. SOX9, AMH, FOXL2 and FST showed the greatest difference between testes and ovaries. WNT4 was significantly higher in females than males over the entire period examined, but was not significantly higher at any given time point.

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Additional file 2:

Raw expression data for qPCR analysis of the oestrogen-treated and control culture gonads. Graphs show expression relative to beta-actin (house keeping control gene) that shows high levels of expression. Small bars represent a small difference between target gene expression and that of beta-actin (thus, represent high expression values) while large bars show a greater difference between target gene levels and beta-actin (thus, representing low expression values). Error bars show one standard deviation either side of the mean.

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Additional file 3:

SOX9 distribution in the bipotential gonad. SOX9 protein was examined in the day 25 fetal XY gonad to determine its distribution at the start of the culture period. At this time, SOX9 was already localized in the nuclei (black arrowhead) of many somatic cells, but some weak cytoplasmic staining (red arrowhead) also remained.

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Additional file 4:

Primers used. Primers used in quantitative PCR reactions are listed in 5' to 3' orientation.

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Additional file 5:

Raw expression data for qPCR analysis of the normal gene profiles shown in Figure 1. Mean delta Ct values and the standard deviation (std-dev) for each data point shown in Figure 1. The housekeeping gene β-actin shows high levels of expression so most delta Ct values are negative. The left column represents data points from the male profile and the right column represents data points from the female profile. Gene names are indicated in the top left corner of each table. Graphed data is log transformed and shown relative to beta actin in Figure 1.

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