Plots of expression level in log scale versus four time points for the 3,890 diurnally regulated transcripts arranged in eight clusters

<p><b>Copyright information:</b></p><p>Taken from "Genome-wide expression profiling and bioinformatics analysis of diurnally regulated genes in the mouse prefrontal cortex"</p><p>http://genomebiology.com/2007/8/11/R247</p><p>Genome Biology 2007;8(11):R247-R247.</p><p>Published online 20 Nov 2007</p><p>PMCID:PMC2258187.</p><p></p> These clusters have very distinct temporal patterns of expression variation, suggesting that the clustering procedure is effective in picking out signals specific to each cluster

Heat map of enriched TFBSs and their corresponding TFs for each of the eight clusters, when both TFBSs and TFs are present in the diurnal genes

<p><b>Copyright information:</b></p><p>Taken from "Genome-wide expression profiling and bioinformatics analysis of diurnally regulated genes in the mouse prefrontal cortex"</p><p>http://genomebiology.com/2007/8/11/R247</p><p>Genome Biology 2007;8(11):R247-R247.</p><p>Published online 20 Nov 2007</p><p>PMCID:PMC2258187.</p><p></p> The columns indicate the TFs in each of the eight clusters, where the rows represent the enriched (< 0.05) TFBSs in the 1 kb upstream region of genes in each of the eight clusters. The color of the cell represents the degree of matching: green cells indicate that there is no matching TF and TFBS, while increasing intensity of red colors indicate one or more matches. We found that clusters 2, 7 and 8 contain few TFs that may regulate genes in other clusters, but clusters 4-6 tend to have TFs that may regulate genes in most of the other clusters

VEGF-Mediated Proliferation of Human Adipose Tissue-Derived Stem Cells

<div><p>Human adipose tissue-derived stem cells (ADSCs) are an attractive multipotent stem cell source with therapeutic applicability across diverse fields for the repair and regeneration of acute and chronically damaged tissues. In recent years, there has been increasing interest in ADSC for tissue engineering applications. However, the mechanisms underlying the regulation of ADSC proliferation are not fully understood. Here we show that 47 transcripts are up-regulated while 23 are down-regulated in ADSC compared to terminally differentiated cells based on global mRNA profiling and microRNA profiling. Among the up-regulated genes, the expression of vascular endothelial growth factor (VEGF) is fine-tuned by miR-199a-5p. Further investigation indicates that VEGF accelerates ADSC proliferation whereas the multipotency of ADSC remains stable in terms of adipogenic, chondrogenic and osteogenic potentials after VEGF treatment, suggesting that VEGF may serve as an excellent supplement for accelerating ADSC proliferation during <i>in vitro</i> expansion.</p></div

MiR-199a-5p-mediated VEGF down-regulation in ADSCs.

<p>(A) Targetscan indicates that miR-199a-5p binds the 3′UTR of VEGF mRNA; (B) Lentiviral infection efficiency was evaluated by ZsGreen intensity; (C) Dual-luciferase reporter assay indicated that miR-199a-5p interacted with the 3′UTR of VEGF mRNA by using lentiviral infection (n = 3); (D) qRT-PCR assay indicated that the lentiviruses overexpressed miR-199a-5p (n = 3); (E) qRT-PCR assay indicated that miR-199a-5p overexpression in ADSCs reduced VEGF mRNA levels by approximately 42% (n = 3). (F) ELISA assay indicated that miR-199a-5p overexpression in ADSCs reduced VEGF protein levels by approximately 16% (n = 5). The values were mean ± SEM (n = 5). **<i>P</i><0.01.</p

Global expression profiles of mRNAs and microRNAs.

<p>(A) Hot map of mRNA levels. (B) Hot map of miRNA levels. A set of mRNAs and miRNA were significantly differentially expressed in human endothelial cells, human fibrobast cells and ADSCs. Red color indicated high expression whereas yellow color indicated low expression. H1, human endothelial cell; S1, human fibroblast cell; E1–E6, human adipose tissue-derived stem cells (ADSCs).</p

VEGF-mediated proliferation of ADSCs.

<p>(A) MTT assay indicated that proliferation of ADSCs was accelerated by supplementation of a gradient concentration of VEGF (n = 5); (B) Trypan blue staining confirmed that the effect of VEGF on ADSC proliferation was dose-dependent (n = 3, p<0.01); (C) ELISA assay indicated that VEGF expression was reduced by approximately 40% in ADSCs after shRNA silencing (n = 5, p<0.01); (D) Cell number was significantly reduced when ADSCs were cultured in FBS-containing medium for 24 or 48 hours after VEGF silencing (n = 3, P<0.01); (D) The cell number reduction was also confirmed when ADSCs were cultured for 24 or 48 hours in KSR-containing medium after VEGF silencing (n = 3, P<0.01); (E) ADSC proliferation was also evaluated in FBS medium by trypan blue assay when VEGF expression was knocked down, it was demonstrated that VEGF reduction resulted in inhibition of ADSC proliferation (n = 5). * <i>P</i><0.05, **<i>P</i><0.01.</p

Down-regulated microRNAs.

<p>Down-regulated microRNAs.</p

The maintenance of multipotent cell surface markers in ADSCs.

<p>(A) CD29 and CD44 staining revealed that cells had a similar pattern of expression compared to the untreated group after VEGF treatment; (B) Flow cytometry indicated that ADSC surface markers (CD29 and CD44) showed similar abundance compared with the untreated group.</p

Up-regulated microRNAs.

<p>Up-regulated microRNAs.</p

Up-regulated and down-regulated genes in both arrays.

<p>P Value<0.01, fold change >2 or fold change <0.5.</p