Open Access Research article

Novel polysome messages and changes in translational activity appear after induction of adipogenesis in 3T3-L1 cells

Carolin Fromm-Dornieden1, Silvia von der Heyde2, Oleksandr Lytovchenko1, Gabriela Salinas-Riester3, Bertram Brenig1, Tim Beissbarth2 and Bernhard G Baumgartner4*

Author Affiliations

1 Institute of Veterinary Medicine, University of Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany

2 Statistical Bioinformatics, Department of Medical Statistics, University Medical Center, Humboldtallee 32, 37073 Göttingen, Germany

3 DNA Microarray Facility Göttingen, Department of Developmental Biochemistry, University of Göttingen, Humboldtallee 23, 37073 Göttingen, Germany

4 Department of Internal Medicine, Metabolic Diseases and Medical Molecular Biology, Paracelsus Private Medical University Salzburg, Müllner Hauptstr. 48, 5020 Salzburg, Austria

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BMC Molecular Biology 2012, 13:9  doi:10.1186/1471-2199-13-9

Published: 21 March 2012

Additional files

Additional file 1:

Microscopical control of adipogenesis. In comparison to the fibroblastic phenotype of 3T3-L1 preadipocytes (left picture), mature adipocytes´ phenotype (right picture) is round and cells accumulate lipid droplets in the cytoplasm. 3T3-L1 cells two days before hormonal induction (left picture) were stained with Coomassie blue. Cells nine days after hormonal induction (right picture) were stained with Oil Red O (400× magnification).

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Additional file 2:

Molecular control of adipogenesis. Analysis of mRNA steady state levels of C/EBPβ (empty boxes) and PPARγ (filled boxes) by means of q-PCR in total RNA at time points 0, 0 + 6 h and 9 days. Ct-values were calibrated to day 0, normalized with βActin, (mean of 3 experiments with 3 replicates each, n = 9). C/EBPβ was up-regulated 3times at 0 + 6 h and back to base levels at day 9. PPARγ was up-regulated at day 9, no significant change of mRNA steady state levels were detected at day 0 + 6 h. Standard deviations are shown by error bars.

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Additional file 3:

Control of stability of gradients. Stability of 14 linear gradients was proved with a refractometer. Gradient fractions were collected from top of gradient and percentage of sucrose content was measured. Standard deviations are shown by error bars.

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Additional file 4:

Sucrose gradient analysis. Polysomal RNA (28S/18S ratio ~ 2) was separated from non-polysomal RNA (28S/18S ratio ≠2) by sucrose gradient centrifugation. Ratio of 18S and 28S rRNA was measured to obtain the polysome profile on Agilent 2100 Bioanalyzer. Fractions 5 to 7 contain non-polysomal RNA and fractions 9 to 11 polysomal RNA.

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Additional file 5:

Further information to Table 1. Gene functions, GenBank accession numbers and fold change for mRNAs that are fourfold and greater up- or down-regulated 6 hours after stimulation of adipogenesis.

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Additional file 6:

Heatmap forhousekeeper-normalized PCR data. Column 1 to 4 shows housekeeper-normalized values for polysomal (p) and non-polysomal (np) fractions at two time points (0 h and 6 h after hormonal induction). Column 5 shows fraction to time ratio (log; (p6-np6) - (p0-np0)).

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Additional file 7:

Results of Cluster analysis by PANTHER DB. This Table contains the name of the PANTHER classification category, the genes that map to the respective category, the expected number of genes in the respective category based on the reference genome, plus or minus signs indicating over- or under-representation of the respective category in the experiment and finally the p-values determined by the binomial statistic according to [60].

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