Multivariate statistical analysis of the MR spectra.

<p>Scores plots (PC1 vs PC2) obtained by performing PCA on the MR spectra acquired on polar extracts (8 replicates per treatment condition) of (A) both PC3 and LNCaP cells, and individual (B) PC3 and (C) LNCaP prostate cancer cells following a 48-hrs treatment with DMSO (solvent control, black), LY294002 (green) and 17AAG (red).</p

Common metabolic changes in prostate and breast cancer cells following drug treatment.

<p>Quantification of selected metabolites (shown as percent of control, mean ± standard deviation) from MR spectra acquired on prostate (PC3 and LNCaP, N = 8) and breast (MCF-7, N = 3) cancer cell lines following 48 hours of treatment with (A) LY294002 or (B) 17AAG. *: p<0.05; **: p<0.005; ***: p<0.0005. Pcholine: phosphocholine.</p

Accumulation of citrate following treatment with 17AAG in PC3 prostate cancer cells.

<p>Enlarged section (2.49 – 2.72 ppm) of the MR spectra acquired on polar extracts of PC3 cells following 48 hours of treatment with DMSO (solvent control, black), LY294002 (green) and 17AAG (red). Spectra were normalized according to the probabilistic quotient normalization method.</p

Multivariate statistical analysis of the MR spectra.

<p>Scores plots (PC1 vs PC2) obtained by performing PCA on the MR spectra acquired on polar extracts (8 replicates per treatment condition) of PC3 and LNCaP prostate cancer cells. Control samples were compared to samples treated for 48 hours with either LY294002 ((A) for PC3 and (C) for LNCaP cells) or 17AAG ((B) for PC3 and (D) for LNCaP cells).</p

Multivariate statistical analysis of the MR spectra.

<p>Loadings plots (on PC1) obtained by performing the PCA comparisons on the MR spectra of control and one treatment per analysis (as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0026155#pone-0026155-g003" target="_blank">Fig. 3</a>) acquired on polar extracts of PC3 (black line) and LNCaP (red line) prostate cancer cells following 48 hours of treatment with (A) LY294002 or (B) 17AAG. Enlarged sections of the loadings plots represent the region of 1.9–4.1 ppm. Ala: alanine; Asn: asparagine; Cho: choline; Cit: citrate; Cre: creatine; Fum: fumarate; Glc: glucose; Gln: glutamine; Gly: glycine; GPcho: glycerophosphocholine; GSH: glutathione; His: histidine; Ile: isoleucine; Lac: lactate; Leu: leucine; m-Ino: myo-inositol; Pcho: phosphocholine; Pcre: phosphocreatine; Phe: phenylalanine; Tau: taurine; Tyr: tyrosine; Val: valine, UDPS: UDP sugars.</p

Inhibition of target signaling pathways in cancer cells following drug treatment.

<p>Schematic of signaling pathways targeted by LY294002 and 17AAG and Western blots showing modulation of p-4E-BP1 and c-Raf (β-actin as loading control) levels following administration of DMSO (solvent control; C), LY294002 (L) or 17AAG (A).</p

PKM1 and PKM2 protein expression in a series of WHO grade I-IV human astrocytoma specimens.

<p><b>A</b>, Protein isolated from frozen normal brain (NB), grade (Gr) I-IV astrocytoma, and human GBM cell lines (U87, G55, U251) was subjected to Western blot analysis using PKM1-, PKM2-, and β-actin-specific antibodies. <b>B</b>, PKM signal intensity values derived from A and normalized to β-actin. <b>C</b>, IHC analysis of representative fixed sections from tumors in panel A using PKM1- or PKM2-specific antibodies. For Gr-IV sections T = tumor, N = tumor-infiltrated normal brain. <b>D</b>, Pyruvate kinase activity (mean±SE) determined using an LDH-coupled enzyme assay, N = 5 for each group.</p

Quantification of CTP:phosphocholine cytidylyltransferase activities.

<p>(A) Buildup of CDP-choline peaks in MCF7 cell lysates over 1.5 hours; (B) Time courses of CDP-choline production in control and SAHA-treated cells, showing no significant change in cytidylyltransferase activity with treatment.</p

Detection of endogenous metabolites by magnetic resonance spectroscopy.

<p>(A) Representative ³¹P spectra of control (bottom) and SAHA-treated (top) MCF7 cell extracts, showing increases in PC and GPC after 48-hour treatment; (B) Representative ¹H spectra (3.10–3.30 ppm) of choline-containing metabolites, highlighting increased tCho levels after 48-hour treatment.</p

Immunoblots probing the protein expression of PLA2G4C, ChoKα SLC44A1.

<p>β-actin served a loading control.</p