Effect of aerobic exercise on intramyocellular (IMCL) and intrahepatocellular lipids (IHCL).

<p><b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070865#pone-0070865-g003" target="_blank"><b>Figure 3a</b></a></b> Individual IMCL and IHCL measurements before (pre) and after (post) a 2h-aerobic exercise at 50% VO_2max of each subject. A significant decrease in IMCL (p<0.005) and a significant increase in IHCL (p<0.002) was documented. The diagram on the far right is a blow up of the IHCL data depicted in the middle part to show the changes in IHCL for volunteers with low IHCL levels. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070865#pone-0070865-g003" target="_blank"><b>Figure 3b</b></a> Changes (mean ± 1 SEM) of IMCL and IHCL (in % from baseline) following a 2h-aerobic exercise at 50% VO_2max.</p

Study protocol.

<p>The clinical protocol included three visits and two periods. Period 1: weight maintaining diet, period 2: no physical activity, high fat diet (as additional fat snacks, 0.75 g fat/kgBW for 1.5 days prior to visit 3). Visit 1 Spiroergometry (calculation of 50% VO_2max, trial on the treadmill). Visit 2 Two-step hyperinsulinaemic euglycaemic clamp using stable isotope technique. Visit 3 Assessment of intramyocellular (IMCL) and intrahepatocellular lipids (IHCL) before and after two hours aerobic exercise at 50% VO_2max. Measurements of subcutaneous and visceral fat mass using MRI imaging.</p

Clinical parameters, body composition and fat availability, exercise parameters, insulin sensitivity and MR-spectroscopy data in endurance trained athletes.

<p>Values are indicated in mean ± SEM. BMI  =  body mass index; FFA  =  free fatty acids; suppression of EGP  =  suppression of endogenous glucose production  =  measure of hepatic insulin resistance; M-value  =  glucose infusion at high insulin dose  =  measure of peripheral (whole body) insulin resistance; IMCL  =  intramyocellular lipids; IHCL  =  intrahepatocellular lipids expressed as % of hepatic water signal; IMCL pre/IHCL pre  =  before exercise; IMCL post/IHCL post  =  post exercise; ΔIMCL/ΔIHCL  = 2h-aerobic exercise induced changes of IMCL/IHCL; α =  p-value <0.002; β  =  p value <0.001.</p

Effect of aerobic exercise on serum free fatty acid concentrations.

<p>Individual (left) and overall (mean ± 1 SEM) levels of circulating FFA during the course of the 2 h-aerobic exercise at 50% VO_2max.</p

Flow sheet of patients, intervention and analysis.

<p>Flow sheet of enrolled patients, intervention and analysis of data. IMCL  =  intramyocellular lipids.</p

<i>ELF5</i> expression in normal breast and breast cancer.

<p><i>ELF5</i> expression in the UNC337 breast cancer cohort according to subtype. Top panel shows combined results with statistical analysis (thick black line, median;, box, interquartile range 25%–75% of points; whiskers, 1.5× interquartile range; crosses, individual tumors). Bottom panel <i>ELF5</i> expression in all individual tumor and normal samples. Basal and normal had significantly higher expression of ELF5.</p

ELF5 suppresses the estrogen-sensitive phenotype.

<p>(A) Western blot showing reduced expression of key genes involved in the response to estrogens following induction of ELF5 expression. (B) Reduced transcriptional activity of reporters of <i>ER</i> and <i>FOXA1</i> (<i>UGT2B17</i> promoter) transcriptional activity following induction of ELF5 in MCF7 cells. Black bars, -DOX, grey bars +DOX 72 h for <i>ERE</i> and 24 h and 48 h for <i>FOXA1</i>. (C) Cell accumulation in MCF7-V5 cell cultures with (+E) or without (−E) 10 nM estrogen treatment, or following expression of ER (+ER) and 10 nM E in the context of induced ELF5. Black bars, -DOX; grey bars +DOX, 72 h and 144 h, respectively. (D) interaction of ELF5-regulated gene sets with estrogen-regulated gene sets. <i>p</i>-Values and odds ratios derived from hypergeometric tests. Number of genes in brackets. (E) Enrichment of gene sets in ELF5 ChIP targets either down (Dn) or Up in response to forced ELF5-V5 expression in T47D cells with DOX. P-Values for hypergeometric tests from GSEA (upper case) or Oncomine (lower case).</p

Elf5 modulates the adhesion of breast cancer cells.

<p>(A) Quantification of detached cells in cultures treated with DOX (+D) compared to no induction (−D). (B) Ability of DOX-treated cells to replate 4 h after trypsin destruction of attachment proteins, compared to untreated cells. Data are expressed as a percentage of replated untreated cells. (C) Proportion of apoptotic cells in DOX treated (grey bars) compared to untreated (black bars) T47D-ELF5-V5 cells, measured by flow cytometry using the M30 antibody. (D) Expression and activation of key cell adhesion proteins following DOX induction of ELF5-V5 expression.</p

ELF5 specifies breast cancer subtype.

<p>(A) Sub network of breast cancer subtype gene sets derived from forced ELF5 expression in MCF7 luminal breast cancer cells (inner node color) and knockdown of ELF5 expression in HCC1937 basal breast cancer cells. Node size is proportional to gene set size; thicker green lines indicate greater gene set overlap. Nodes are positioned according to similarity in gene sets. Labels in bold type indicate the functional significance of the four clusters generated, label is plain type is the gene set name. The full network is shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001461#pbio.1001461.s016" target="_blank">Figure S16</a>. (B–D) expression signature analysis of the ELF5-induced changes in molecular subtype produced by ELF5 knockdown in HCC1937 cells (B), or forced ELF5 expression in MCF7 cells (C), or T47D cells (D). Bars show the indicated comparisons that produce the associated <i>p</i>-values. BS, borderline significance; NS, not significant.</p

Visualization of the transcriptional functions of ELF5 in breast cancer.

<p>Affymetrix transcript profiling following induction of ELF5 in T47D and MCF7 luminal breast cancer cells, analysed by LIMMA and GSEA. <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001461#s2" target="_blank">Results</a> are visualized using the enrichment map plug-in for Cytoscape. Each circular node is a gene set with diameter proportional to the number of genes. The outer node color represents the magnitude and direction of enrichment (see scale) in T47D cells, inner node color enrichment in MCF7 cells. Thickness of the edges (green lines) is proportional to the similarity of gene sets between linked nodes. The most related clusters are placed nearest to each other. The functions of prominent clusters are shown. The network can be examined in detail using the scalable PDF in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001461#pbio.1001461.s004" target="_blank">Figure S4</a>.</p