Open Access Highly Accessed Research article

Expression profiling of genes regulated by TGF-beta: Differential regulation in normal and tumour cells

Prathibha Ranganathan1, Animesh Agrawal1, Raghu Bhushan1, Aravinda K Chavalmane1, Ravi Kiran Reddy Kalathur1, Takashi Takahashi2 and Paturu Kondaiah1*

Author Affiliations

1 Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560012, India

2 Division of Molecular Carcinogenesis, Centre for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Japan

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BMC Genomics 2007, 8:98  doi:10.1186/1471-2164-8-98

Published: 11 April 2007

Additional files

Additional file 1:

List of genes commonly regulated by TGF-β in HPL1D and A549 cell-lines. A list of genes, which are regulated by TGF-β in both HPL1D and A549 cell-lines along with the fold changes at 1, 4 and 12 hrs of treatment.

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Additional file 2:

List of genes regulated by TGF-β in only A549 cell-line. A list of genes, which are regulated by TGF-β in A549 alone along with the fold changes at 1, 4 and 12 hrs of treatment.

Format: XLS Size: 469KB Download file

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Additional file 3:

List of genes regulated by TGF-β only in HPL1D cell-line. A list of genes, which are regulated by TGF-β in HPL1D cell-line alone along with the fold changes at 1, 4 and 12 hrs of treatment.

Format: XLS Size: 206KB Download file

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Additional file 4:

Semi-quantitative RT-PCR analyses of selected genes with respect to regulation by TGF-β in A549 and HPL1D cells. The respective cell-lines were grown to 90% confluence (as described in methods section), washed with serum free medium, treated with 5 ng/ml TGF-β 1 for 1, 4, 6, 12 and 24 hours. Each treatment also has untreated cells as control at each time-point. Two microgram of total RNA from each treatment was reverse transcribed and cDNA equivalent to 20 ng total RNA was used for the PCR reactions. All PCR reactions were done under non saturating conditions. The products were resolved on 2% agarose gel and the gel pictures were taken on Kodak Image station 440CF. The band intensities were quantified using Kodak 1D 3.6 software. A, ethidium bromide staining pattern of the PCR products. B and C graphs representing the fold change over untreated controls after normalization with the expression of RPL35a, in HPL1D and A549 cells respectively.

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Additional file 5:

Induction of pSBE-Luc activity by TGF-β in A549 cells. For SBE-luc induction by TGF-β in A549 cells, twenty five thousand cells were plated in 24 well dishes 16–24 hours prior to transfection. 200 ng of pSBE-luc plasmid and 1.25 ng of pRL-CMV construct (Renilla luciferase, for transfection normalization) were transfected using Effectene reagent (Qiagen GmbH, Germany) in serum free conditions for 12 hrs. The cells were recovered in medium containing 10% FBS for 24 hours, washed with serum free medium for 24 hours (3 changes) and then treated with 5 ng/ml TGF-β for 18 hours. The cells were then lysed and lysates were used for dual-luciferase assay (Promega inc, USA). The ratio of the firefly-luciferase to renilla-luciferase has been plotted on the y-axis.

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