Dynamic signal evolution across (N = 7) injections.

<p>The mean signal for lactate and pyruvate, normalized to peak carbon signal for each injection, are displayed with error bars that indicate the minimum and maximum values at each time over all injections. Total HP ¹³C was estimated by summing signal from HP ¹³C Lactate and HP ¹³C Pyruvate. The average linewidth for pyruvate and lactate peaks were 19±5 Hz and 17±5 Hz, respectively.</p

Spectroscopic images of the reaction carried out in a standard imaging phantom.

<p>Proton imaging (top left) shows the phantom structure in high resolution. Spectroscopic imaging data acquired using a radial EPSI sequence allows metabolite-specific visualization of tracer distribution (bottom row). Spectroscopic data can be intrinsically registered to high-resolution proton images (top center and right).</p

A schematic view of the dynamic chemical phantom structure.

<p>The injection and exhaust ports were fitted with catheters to facilitate rapid mixture of reagents at isocenter. A thin acrylic sheet was attached to the top to seal the fill cavity. This top could be removed to allow cleaning after injection. The phantom rested on a sled that allowed convenient removal and insertion of the phantom and included warm circulating water to maintain constant temperature.</p

The mean, standard deviation and coefficient of variation for all repetitions (N = 7) of the dynamic phantom.

<p>Lac and Pyr refer to the total volume under the spectral and temporal curves for each tracer; <i>k_PL</i> is the forward reaction rate (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071274#pone.0071274.e006" target="_blank">Equations 6</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071274#pone.0071274.e010" target="_blank">8</a>).</p

Additional file 1: Figure S1. of Induction of autophagy by ARHI (DIRAS3) alters fundamental metabolic pathways in ovarian cancer models

Growth of parental and ARHI-transfected SKOv3 and Hey cells. Effect of Dox treatment on the growth of parental and ARHI-transfected SKOv3 and Hey cells at 24 and 48 h. Western analysis of the effect of ARHI expression on LC3I and LC3II is also presented. Figure S2. Western analysis of GLUT1 expression following ARHI induction. Figure S3. Analysis of ARHI expression and autophagy markers during Atg5 knockdown. Effect of Atg5 knockdown on LC3I and LC3II levels during ARHI expression in SKOv3-ARHI cells. Immunofluorescence of SKOv3-ARHI cells transfected with GFP-LC3 following ARHI induction with and without Atg5 knockdown. Figure S4. Western analysis of LDH and CK expression following ARHI induction. Figure S5. Induction of ARHI expression in vivo. Expression of ARHI and LC3 in subcutaneous SKOv3-ARHI tumors at 24-72 h post-treatment with Dox. Figure S6. Expression of ACC and Phsopho-ACC by RPPA. Figure S7. Fractional 13C label incorporation from 5-13C-Gln in SKOv3-ARHI. The fractional incorporation of the glutamine 13C label into NMR-observable intracellular metabolites following induction of ARHI. (PDF 867 kb