Distribution levels in brain choroid plexus or spleen of mice intraperitoneally injected with HPMCs.

<p>Representative GPC profiles of fluorescein-labeled TC-5RW are shown for brain choroid plexus (A) and spleen (B) of ddY mice at 1 week after intraperitoneal injection. The distribution levels of fluorescein-labeled TC-5RW are shown for brain choroid plexus (C, E) and spleen (D, F) of ddY, ICR, and Tg7 mice at 1 week (C, D) or 8 weeks (E, F) after intraperitoneal injection. It is noted that there is no significant difference among the three mouse strains.</p

Relationship of the distribution level with the anti-prion activity.

<p>The distribution levels of fluorescein-labeled HPMCs in brain choroid plexus (A, C) or spleen (B, D) of ddY mice are plotted against the anti-prion activities of the immediate post-infection (A, B) or of the 1-year pre-infection (C, D) treatments, which are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185357#pone.0185357.t001" target="_blank">Table 1</a>.</p

Chemical and biological properties of HPMCs.

<p>Chemical and biological properties of HPMCs.</p

Thermal characteristics of HPMCs.

<p>Thermal characteristics of HPMCs.</p

Relationship between the distribution and the molecular weight of HPMCs.

<p>The distributions in brain choroid plexus (A) or spleen (B) are plotted against the <i>M</i>_n of HPMCs in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185357#pone.0185357.t001" target="_blank">Table 1</a>.</p

Relationships of the pyrene index with the macrophage uptake level of HPMCs.

<p>The data of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185357#pone.0185357.g004" target="_blank">Fig 4B</a> are compared with those of the macrophage uptake level, which are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185357#pone.0185357.t001" target="_blank">Table 1</a>.</p

Representative thermal profile of TC-5RW.

<p>(→) and (←) indicate heating and cooling processes, respectively.</p

FTIR spectra of the fingerprint region of HPMCs.

<p>IR spectra were superimposed.</p

CE effects on PrP expression and PrP^Sc formation.

<p>(a) Immunoblotting of PrP^C in the brain and spleen from Tg7 mice treated with TC-5RW (4 g/kg body weight) or vehicle at the designated time points (<i>n</i> = 3 for each time point). Molecular size markers on the right indicate sizes in kDa. The mean and standard deviation are shown in each bar graph. (b) PrP mRNA expression level in the brain and spleen from Tg7 mice treated with TC-5RW (4 g/kg body weight) or vehicle at designated time points. The mean and standard deviation (<i>n</i> = 3 for each time point) are shown. (c) Immunoblotting of protease-resistant PrP^Sc in the protein misfolding cyclic amplification reaction performed in the presence of TC-5RW. Representative quadruplicate data are shown for the cyclic amplification of 263K prion performed in the presence (+) or absence (−) of TC-5RW or hydroxypropyl methyl dextran-70 kDa (HP-M-dextran-70) at a dose of 10 μg/mL. HP-M-dextran-70 was used as a negative control. In the absence of each compound, the vehicle, water, was added to the reaction mixture. (d) Immunoblotting of protease-resistant PrP^Sc in RML prion-infected N2a cells treated with HPMCs. Signals for β-actin are shown as controls for the integrity of samples used for PrP^Sc detection. The chemical property and <i>in vivo</i> efficacy of TC-5EW are shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006045#ppat.1006045.s004" target="_blank">S4 Fig</a>. (e) Immunoblotting of PrP^C in uninfected N2a cells treated with HPMCs at a dose of 1 mg/mL in duplicate. (f) Flow cytometry of cell surface PrP^C in uninfected N2a cells treated with HPMCs at a dose of 1 mg/mL. Black and blue lines indicate vehicle-treated cells and HPMC-treated cells, respectively. The broken line peaks on the left show the respective isotype controls.</p

Macrophage uptake analysis.

<p>(a) HPMC macrophage uptake ratio. This represents the percentage of the HPMC amount of macrophage lysate <i>vs</i>. the amount added into the culture medium. The mean and standard deviation of triplicate experiments are shown. (b) Analysis of the linear correlation between the macrophage uptake ratio and prophylactic efficacy of HPMCs. Survival times of Tg7 mice treated with HPMCs immediately post-infection or treated 12 months prior to infection were obtained from Figs <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006045#ppat.1006045.g001" target="_blank">1c</a> and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006045#ppat.1006045.g004" target="_blank">4a</a>. Statistical correlations were identified using the Pearson’s correlation coefficient.</p