HCV E2 inhibited stabilization of AIMP1/p43 by grp78.

<p>A. Plasmid expressing grp78 was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. β-tubulin was used as a loading control. EV; empty vector. B. grp78 siRNA or negative control siRNA was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. C. grp7 siRNA was transfected to Huh-7 cells at indicated amounts. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody.</p

HCV E2 induced ubiquitin-mediated degradation of AIMP1/p43.

<p>A. Plasmid expressing HCV E2 was transfected to Huh-7 cells. Medium was changed 6 hours after transfection and MG132 was added at the concentration of 50 µM in 20 hours. Western blot analysis was done 6 hours after the treatment. β-tubulin was used as a loading control. EV; empty vector. B. Plasmid expressing HA-tagged ubiquitin and plasmid expressing HCV E2 were transfected to Huh-7 cells. Medium was changed 6 hours after transfection and MG132 was added at the concentration of 50 µM in 20 hours. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody and protein G.</p

HCV E2 and cellular AIMP1/p43 interacted with each other.

<p>A. GST pulldown assay showed interaction of the two proteins. Purified GST-E2 was used for binding to expressed AIMP1/p43 in HEK 293 cells. Purified GST was used as a control. B. V5-tagged E2 was expressed in HEK 293 cells and immunoprecipitation was done with anti-V5 antibody. IgG was used as a control. C. C-terminal V5-tagged core-E1 or core-E1-E2 were expressed in HEK 293 cells and immunoprecipitation was done with anti-AIMP1/p43 antibody. D. The two proteins showed subcellular colocalization. Plasmid expressing AIMP1/p43 and plasmid expressing V5-E2 were transfected to Huh-7 cells. 24 hours' after transfection, cells were treated with 10 µM MG132 for 6 hours. Cells were immunostained with anti-AIMP1/p43 antibody (green) and anti-V5 antibody (red) and observed under confocal microscope. E. Native E2 and AIMP1/p43 showed subcellular colocalization. Plasmid expressing AIMP1/p43 and plasmid expressing C-E1-E2-V5 was transfected to Huh-7 cells. 24 hours' after transfection, cells were treated with 10 µM MG132 for 6 hours. Cells were immunostained with anti-AIMP1/p43 antibody (green) and anti-V5 antibody (red) and observed under confocal microscope. F. V5-tagged HCV E2 and AIMP1/p43 was observed in proximity ligation assay (PLA). Red dots show the interaction between E2 (either 2a or 1b type) and AIMP1/p43. Nucleus was stained with DAPI (blue).</p

Endogenous AIMP1/p43 decreased in the presence of HCV E2.

<p>A. Plasmid expressing HCV E2 was transfected to Huh-7 cells and endogenous AIMP1/p43 was detected by anti-AIMP1/p43 antibody. The amount of plasmid used for transfection was 0.5, 1, and 2 µg for each lane. β-tubulin was used as a control. EV; empty vector. B. Plasmid expressing HCV core, HCV E1, HCV E2, HCV NS3, or HCV NS5A was transfected to Huh-7 cells and the change of amount of endogenous AIMP1/p43 was detected by anti-AIMP1/p43 antibody. C. Plasmid expressing HCV core, core-E1, or core-E1-E2 was transfected to Huh-7 cells and the enogenous AIMP1/p43 expression level was observed. D. JFH1 HCVcc was infected into Huh7.5 cells. The expression level of endogenous AIMP1/p43 was detected by anti-AIMP1/p43 and expression of HCV RNA was detected by anti-core antibody. E. Plasmid expressing HCV E2 was transfected to Huh-7 cells and expressed for 24 hours. Total RNA was isolated and real-time RT-PCR was performed. The average of three independent tests is shown with error bars. F. Plasmid expressing FLAG-tagged E2 was transfected to Huh-7 cells. 24 hours' after transfection, immunostaining was done using anti-FLAG antibody (red) and anti-AIMP1/p43 antibody (green). Cells were observed under fluorescencemicroscope.</p

HCV E2 promotes cell surface expression of gp96 and TGF-β signaling.

<p>A. Palsmid expressing HCV E2, NS3, NS5A, or NS5B was transfected to Huh-7 cells and grown for 24 hours. Cell surface gp96 was probed with anti-gp96 antibody in flow cytometry. B. The same experiment with HCV E2 was carried out in Bjab cells. C. JFH1 particle (HCVcc) was infected to Huh7.5 cells. Cell surface gp96 was probed with anti-gp96 antibody using flow cytometry. D. HCV pseudoparticle (HCVpp) was infected to Huh7.5 cells. Cell surface gp96 was probed with anti-gp96 antibody using flow cytometry. E. A reporter plasmid, p3TP-lux, was transfected into Huh-7 cells along with 0.5, 1.0, or 2.0 µg of plasmid expressing HCV E2 and luciferase activity was measured in a luminometer. Transfection efficiency was normalized with cotransfection of pCH110 harboring LacZ gene and measuring β-galactosidase activity. Experiment was done in a triplicate. F. Plasmid expressing HCV E2 (either 0.5 µg or 2 µg) were used to transfect Huh-7 cells and PAI-1 expression was observed. TGF-β was added at the concentration of 4 ng/ml for 20 hours (lanes 4, 5, and 6) and PAI-1 expression was detected using western blotting with anti-PAI-1 antibody. EV; empty vector.</p

HCV E2 inhibited the interaction between grp78 and AIMP1/p43.

<p>A. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. B. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with 2 or 4 µg of plasmid expressing HCV E2 and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. C. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with plasmid expressing HCV E2, E1, core, or NS5A and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody.</p

Beyond “Doing as the Romans Do”: A review of research on countercultural business practices

AbstractThere has long been a dominant logic in the international business literature that multinational corporations should adapt business practices to “fit” host cultures. Business practices that are congruent with local cultural norms have been advocated as effective and desirable, while practices that are incongruent have been deemed problematic. We examine and challenge this persistent assumption by reviewing the literature showing evidence for both benefits and acceptance of countercultural practices (i.e., practices that are seemingly incongruent with local cultural norms or values), and disadvantages and rejection of local practices. Drawing on the literature reviewed, we offer four types of theoretical (ontological, epistemological, causal, and functional) explanations as to why and when countercultural business practices might be preferred. Finally, we provide a springboard for a future research agenda on countercultural practices, centered around understanding the circumstances under which businesses and local stakeholders might benefit from the use of countercultural practices based on such factors as strategic intent, local preferences, institutional drivers, and social responsibility

Additional file 1: Figure S1. of Outcomes of uterine sarcoma found incidentally after uterus-preserving surgery for presumed benign disease

Progression-free survival of patients with unexpected sarcoma stratified by morcellation procedure. Figure S2. Progression-free survival of patients with unexpected sarcoma stratified by morcellation procedure in myomectomy/subtotal hysterectomy group. Figure S3. Progression-free survival of patients with unexpected sarcoma stratified by type of initial surgery in myomectomy/subtotal hysterectomy group. (PPTX 70 kb

Outline of analysis procedures with each geneset showing the general steps required to identify genes that modulate a specific phenotype: selection of genes with the desired phenotype, and identification of phenotype-inducing ARSN and corresponding cancer-associated druggable target genes.

<p>Outline of analysis procedures with each geneset showing the general steps required to identify genes that modulate a specific phenotype: selection of genes with the desired phenotype, and identification of phenotype-inducing ARSN and corresponding cancer-associated druggable target genes.</p

ARSN biology-dominant groups in patients with GBM.

<p>(<b>a</b>) We identified probe sets whose expression most strongly correlated with survival (Kaplan-Meier plots versus survival times, log-rank t-test <0.05). This analysis identified that 122 resulting probe sets of ARSN, DTGs, and PPIs that were correlated with survival in patients with GBM. Then, we performed a supervised clustering with the probesets and GBM subtypes such as proneural (PN), proliferative (Prolif) and mesenchymal (Mes). This analysis showed that 61 probeset as signature genes were differentially expressed in the three discrete subgroups. The 61 probe sets are presented in matrix format, where rows represent individual genes and columns represent each tissue. Each cell in the matrix represents the expression level of a gene in an individual tissue. Red and green cells reflect high and low expression levels, respectively. (<b>b</b>) Tumor subgroups are distinguished by CARS and FARS. Horizontal bars denote mean values. CARS is enriched in Mes and Prolif subgroups, while FARS in PN subgroup. Each Kaplan-Meier plot of overall survival in 130 GBM patients grouped on the basis of expression of CARS and FARS. The difference between two groups was significant when the P value was less than 0.05. (<b>c</b>) Hierarchical clustering of the GSE4290 dataset of 81 GBM samples from patients with GBM and 23 non-tumor tissues based on the 61 probe sets. Each gene with an expression status were shown in Supplementary <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040960#pone.0040960.s021" target="_blank">Figure S21</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040960#pone.0040960.s023" target="_blank">S23</a>. Nine probes were significantly overexpressed in the non-tumor samples, with 2 probes not showing in this analysis.</p