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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Ranganathan et al. BMC Genomics 2007 8:98 doi:10.1186/1471-2164-8-98