Additional file 2: Figure S1. of Epithelial-mesenchymal transition induction is associated with augmented glucose uptake and lactate production in pancreatic ductal adenocarcinoma

Wound healing assay images for Fig. 1f. (DOCX 447 kb

Additional file 1: S1. of Fast exchange fluxes around the pyruvate node: a leaky cell model to explain the gain and loss of unlabelled and labelled metabolites in a tracer experiment

Baseline enrichment of unlabelled experiment and comparison to labelled experiment. S2. GC-MS peaks from DMEM media with and without serum and from HPDE/PDAC cell culture medium. S3. GC-MS chromatogram of low glucose DMEM derivatised by MOX-TMS. S4. Methoximation-acetylation-butylation (MAB) derivatization. S5. Flux analysis for FM and EM experiments. S6. Pathway carbon atom mapping. S7. Intracellular enrichment of PANC-1 cells at 10 hour. S8. OpenFLUX model for HEK 293 and PANC-1 cells. (DOCX 3503 kb

Indirect calorimetry and urinary nitrogen excretion.

a<p>Indirect calorimetry measurements were performed in 9 hr fasted animals following 4–5 weeks of treatment (n = 16/group).</p>b<p>Based on measured urine volume and concentrations of urinary urea and creatinine in a subset of animals (n = 8/treatment group).</p><p>Values are given as means ± SEM.</p><p>Indirect calorimetry and urinary nitrogen excretion.</p

Rates of appearance (<i>R_a</i>) of serine, glycine and arginine in plasma.

<p>Animals per group were n = 6. Values are given as means ± SEM. *P<0.05 versus vehicle.</p

Schematic overview of suggested PPARα mediated changes in amino acid de novo synthesis.

<p>WY 14,643 reduced renal arginine (ARG) output, via down regulation of arginosuccinate synthetase (ASS) and argininosuccinate lyase (ASL), and increased tissue serine (SER) biosynthesis, via up regulation of phosphoserine aminotransferase 1 (PSAT1) with gene regulation previously shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113328#pone.0113328-Sheikh1" target="_blank">[2]</a>. Available hepatic arginine is diverted to polyamine synthesis via altered gene expression including ornithine decarboxylase 1 (ODC1) up regulation <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113328#pone.0113328-Sheikh1" target="_blank">[2]</a>. Serine is used to provide methyl group transfer to support expected PPARα-mediated liver growth and in the process generating glycine (GLY). CIT, citrulline; ORN, ornithine; 3PG, 3-phosphoglycerate.</p

Plasma amino acid concentrations.

<p>Plasma samples were collected after 4 weeks of treatment with WY 14,643 (n = 15) or vehicle (n = 15) in 9 hour fasted rats. ABU, 2-aminobutyric acid. Values are given as means ± SEM. * P<0.05.</p

Hepatic arginase activity measurements in liver homogenates.

<p>Measurements in vehicle and WY 14,643 treated animals (n = 8/group). Values are given as means ± SEM. *P<0.05 versus vehicle.</p

Clinical chemistry data.

<p>Plasma factors in 3 hour fasted rats. TG, triglycerides; FFA, free fatty acids; Rel HOMA_IR (relative to Vehicle Chow diet group HOMA_IR), insulin resistance index. Values are given as means ± SEM with n = 37 per group. #P<0.05 vehicle on fat diet for 1 week versus vehicle on chow diet. *P<0.05 WY 14,643 versus vehicle after 4 weeks of treatment.</p><p>Clinical chemistry data.</p

Estimated energy production from the oxidation of carbohydrate (CHO), lipid and protein, as well as the total energy expenditure (Total).

<p>Measurements were done in the fasting state following 4–5 week vehicle or WY 14,643 treatment in the rats. Values are given as means ± SEM.</p

Body weight development (A) and consumption of high-fat diet (B).

<p>Rats were treated per orally with WY 14,643 (n = 10) or vehicle (n = 10). Values are given as means ± SEM.</p