IBS subtype and CRH-BP SNP.

<p>We analyzed the associations between the SNPs and psychometric scores according to IBS subtype. (a) In individuals with diarrhea-type IBS, <i>rs10474485</i> A allele non-carriers showed higher scores than carriers. There were significant differences in the PSS (p = 0.018) and Trait (p = 0.017) scores. (b) In addition, in the IBS group with mixed symptoms, a significant difference was observed in the SDS score (p = 0.030). There was no significant difference in the constipation group. *p <i><</i> 0.05.</p

Genotype Frequencies in the IBS Patients and Controls.

<p>Genotype Frequencies in the IBS Patients and Controls.</p

Sample Characteristics.

<p>Sample Characteristics.</p

Sex differences in psychometric scores according to the CRH-BP SNP.

<p>(a) In analysis of male subjects (IBS + control), there was no significant difference (SDS, p = 0.864; PSS, p = 0.177; State, p = 0.979; Trait; p = 0.976). (b) In contrast, female A allele non-carriers of <i>rs10474485</i> showed significantly higher PSS (p = 0.028) and State of STAI (p = 0.035) scores and tended to have higher SDS (p = 0.083) and Trait of STAI (p = 0.078) scores than carriers. *p < 0.05.</p

Self-rating Depression Scale and the CRH-BP SNP.

<p>We analyzed associations between the selected SNP (<i>rs10474485</i>) and psychometric scores according to sex. (a) In male subjects, a significant group (IBS/control) × rs10474485 genotype interaction (p = 0.045) was observed. (b) Further, there was a significant group (IBS/control) × <i>rs10474485</i> A allele interaction (p = 0.017). *p <i><</i> 0.05.</p

Psychometric Scores.

<p>Psychometric Scores.</p

The Phospholipid:Diacylglycerol Acyltransferase Lro1 Is Responsible for Hepatitis C Virus Core-Induced Lipid Droplet Formation in a Yeast Model System

<div><p>Chronic infection with the hepatitis C virus frequently induces steatosis, which is a significant risk factor for liver pathogenesis. Steatosis is characterized by the accumulation of lipid droplets in hepatocytes. The structural protein core of the virus induces lipid droplet formation and localizes on the surface of the lipid droplets. However, the precise molecular mechanisms for the core-induced formation of lipid droplets remain elusive. Recently, we showed that the expression of the core protein in yeast as a model system could induce lipid droplet formation. In this study, we probed the cellular factors responsible for the formation of core-induced lipid-droplets in yeast cells. We demonstrated that one of the enzymes responsible for triglyceride synthesis, a phospholipid:diacylglycerol acyltransferase (Lro1), is required for the core-induced lipid droplet formation. While core proteins inhibit Lro1 degradation and alter Lro1 localization, the characteristic localization of Lro1 adjacent to the lipid droplets appeared to be responsible for the core-induced lipid droplet formation. RNA virus genomes have evolved using high mutation rates to maintain their ability to replicate. Our observations suggest a functional relationship between the core protein with hepatocytes and yeast cells. The possible interactions between core proteins and the endoplasmic reticulum membrane affect the mobilization of specific proteins.</p></div

Core-induced LD formation is not mediated by inhibition of Hrd1-dependent Lro1 degradation.

<p>(A) The scheme for sample preparation is the same as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159324#pone.0159324.g004" target="_blank">Fig 4A</a>. (B) The CHX chase western blotting of Lro1-Myc and Dga1-Myc in wild-type and hrd1Δ cells. BY4742 and BY4742 <i>hrd1</i>Δ cells were transformed with the pRS315-Lro1-Myc plasmid and the empty vector pKT10-GAL (Control) or pKT10-GAL-core (Core). pRS315 is a CEN-based plasmid containing <i>LEU2</i> as a selection marker [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159324#pone.0159324.ref024" target="_blank">24</a>], and these vectors each contained their own promoter for <i>LRO1</i> and <i>DGA1</i>. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159324#pone.0159324.s005" target="_blank">S5 Fig</a> for the original data. (C) and (D) The intensity was normalized using the actin intensity. The relative levels of the intensity of wild-type cells (C) and hrd1Δ cells (D) are shown (N = 3). (E) The LD levels in hrd1Δ cells were determined as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159324#pone.0159324.g001" target="_blank">Fig 1</a> legend.</p

Localization of DsRed-core and LDs in wild-type and tunicamycin-treated <i>lro1Δ</i> cells.

<p>(A) Wild-type cells carrying the pKT10-GAL-DsRed-core plasmid were cultured in SRM + Gal medium for 3 h. Cells were stained with BODIPY 493/503. The images of DsRed, BODIPY 493/503, the overlay of DsRed (red) and BODIPY (green), and DIC are shown. The representative images were magnified (right panels). (B) Lro1Δ (BY4742 <i>lro1</i>Δ::<i>HYG</i>) cells carrying the pKT10-GAL-DsRed-core plasmid were cultured in SRM + Gal medium and tunicamycin (4 μg/ml) for 3 h and were stained with BODIPY 493/503. Fluorescent images were taken as described above. Scale bars: 5 μm.</p

Lipid analysis for the core-expressing yeast cells.

<p>(A) TLC analyses of the neutral lipids were performed as described previously [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159324#pone.0159324.ref027" target="_blank">27</a>]. The positions of ergosterol (ERG), sterol ester (SE) and triacylglycerol (TAG) are indicated. Lipids species found in wild-type cells, dga1Δ cells and lro1Δ cells carrying the pKT10-GAL-core plasmid before (raffinose) and after 3 h of culture with galactose (Galactose) compared by TLC analysis. (B) TAG species found in wild-type (black bars), dga1Δ (green bars) and lro1Δ (red bars) cells carrying the pKT10-GAL-core plasmid before (raffinose) and after 3 h of culture with galactose (Galactose) by LC/MS analysis are depicted, and their abundances are compared. The signal intensity unit for the mass spectrometry detector is indicated as arbitrary unit (AU) for quantification of the abundance of specific m/z of the indicated lipid species.</p