Figure 1

<p>Mean numbers (±standard error) of eggs, nymphs, adults or all individuals per plant at two sampling dates of three cohorts of the B biotype <i>Bemisia tabaci</i> that were initiated on healthy, TbCSV-infected, or TYLCCNV-infected tobacco plants. Each plant was inoculated with 5 female and 5 male adult whiteflies.</p

Additional file 1: Table S1. of Higher proliferation of peritumoral endothelial cells to IL-6/sIL-6R than tumoral endothelial cells in hepatocellular carcinoma

Primers used for real time-PCR. (DOC 39 kb

<i>Panax notoginseng</i> saponins (PNS) attenuate Th17 cell differentiation in CIA mice via inhibition of nuclear PKM2-mediated STAT3 phosphorylation

Rheumatoid arthritis (RA) is an autoimmune disease with aberrant Th17 cell differentiation. Panax notoginseng (Burk.) F. H. Chen (Araliaceae) saponins (PNS) have an anti-inflammatory effect and can suppress Th17 cell differentiation. To investigate mechanisms of PNS on Th17 cell differentiation in RA, and the role of pyruvate kinase M2 (PKM2). Naive CD4+T cells were treated with IL-6, IL-23 and TGF-β to induce Th17 cell differentiation. Apart from the Control group, other cells were treated with PNS (5, 10, 20 μg/mL). After the treatment, Th17 cell differentiation, PKM2 expression, and STAT3 phosphorylation were measured via flow cytometry, western blots, or immunofluorescence. PKM2-specific allosteric activator (Tepp-46, 50, 100, 150 μM) and inhibitor (SAICAR, 2, 4, 8 μM) were used to verify the mechanisms. A CIA mouse model was established and divided into control, model, and PNS (100 mg/kg) groups to assess an anti-arthritis effect, Th17 cell differentiation, and PKM2/STAT3 expression. PKM2 expression, dimerization, and nuclear accumulation were upregulated upon Th17 cell differentiation. PNS inhibited the Th17 cells, RORγt expression, IL-17A levels, PKM2 dimerization, and nuclear accumulation and Y705-STAT3 phosphorylation in Th17 cells. Using Tepp-46 (100 μM) and SAICAR (4 μM), we demonstrated that PNS (10 μg/mL) inhibited STAT3 phosphorylation and Th17 cell differentiation by suppressing nuclear PKM2 accumulation. In CIA mice, PNS attenuated CIA symptoms, reduced the number of splenic Th17 cells and nuclear PKM2/STAT3 signaling. PNS inhibited Th17 cell differentiation through the inhibition of nuclear PKM2-mediated STAT3 phosphorylation. PNS may be useful for treating RA.</p

Genomic Insights into the Glutathione S-Transferase Gene Family of Two Rice Planthoppers, <em>Nilaparvata lugens</em> (Stål) and <em>Sogatella furcifera</em> (Horváth) (Hemiptera: Delphacidae)

<div><h3>Background</h3><p>Glutathione S-transferase (GST) genes control crucial traits for the metabolism of various toxins encountered by insects in host plants and the wider environment, including insecticides. The planthoppers <em>Nilaparvata lugens</em> and <em>Sogatella furcifera</em> are serious specialist pests of rice throughout eastern Asia. Their capacity to rapidly adapt to resistant rice varieties and to develop resistance to various insecticides has led to severe outbreaks over the last decade.</p> <h3>Methodology/Principal Findings</h3><p>Using the genome sequence of <em>N. lugens</em>, we identified for the first time the complete GST gene family of a delphacid insect whilst nine GST gene orthologs were identified from the closely related species <em>S. furcifera. Nilaparvata lugens</em> has 11 GST genes belonging to six cytosolic subclasses and a microsomal class, many fewer than seen in other insects with known genomes. Sigma is the largest GST subclass, and the intron–exon pattern deviates significantly from that of other species. Higher GST gene expression in the <em>N. lugens</em> adult migratory form reflects the higher risk of this life stage in encountering the toxins of non-host plants. After exposure to a sub-lethal dose of four insecticides, chlorpyrifos, imidacloprid, buprofezin or beta-cypermethrin, more GST genes were upregulated in <em>S. furcifera</em> than in <em>N. lugens</em>. RNA interference targeting two <em>N. lugens</em> GST genes, <em>NlGSTe1</em> and <em>NlGSTm2,</em> significantly increased the sensitivity of fourth instar nymphs to chlorpyrifos but not to beta-cypermethrin.</p> <h3>Conclusions/Significance</h3><p>This study provides the first elucidation of the nature of the GST gene family in a delphacid species, offering new insights into the evolution of metabolic enzyme genes in insects. Further, the use of RNA interference to identify the GST genes induced by insecticides illustrates likely mechanisms for the tolerance of these insects.</p> </div

Transcription profiles of nine <i>N. lugens</i> GSTs and nine <i>S. furcifera</i> GSTs in third instar-nymphs exposed to sub-lethal concentrations of four different insecticides, Imidacloprid, chlorpyrifos, beta-cypermethrin and buprofezin.

<p>For each time point (6, 12, 24 and 48 hours), transcription levels are expressed as mean fold transcription relative to controls (unexposed nymphs) (CT). Yellow, orange and red indicate significant over-transcription, whilst green, blue and black indicate significant under-transcription (Mann-Whitney test <i>P</i>-value<0.05. Data was converted by SPSS v19.0). White indicates no significant transcription variation.</p

Structure of GST genes in insects was drawn using the genomic coordinates of each GST gene ORF region on its scaffold.

<p>A, Structure of GST genes in <i>N. lugens</i>; B, Comparison of Zeta subclass GST genes from four insects. (<i>Nilaparvata lugens</i> (Nl prefix), <i>Drosophila melanogaster</i> (Dm prefix), <i>Nasonia vitripennis</i> (Nv prefix), <i>Bombyx mori</i> (Bm prefix). Horizontal lines represent the scaffold regions which contain the GST gene. Arrow-head broken lines show the evolvement of exon segments between <i>N. lugens</i> and <i>D. melanogaster.</i></p

Phylogenetic relationships of 145 GST proteins from 12 insect species.

<p><i>Anopheles gambiae</i> (Ag, 27), <i>Apis mellifera</i> (Am, 5), <i>Bombyx mori</i> (Bm, 19), <i>Drosophila melanogaster</i> (Dm, 36), <i>Sogatella furcifera</i> (Sf, 9), <i>Nasonia vitripennis</i> (Nv, 11), <i>Locusta migratoria manilensis</i> (Lm, 10), <i>Acyrthosiphon pisum</i> (Ap, 8), <i>Toxoptera citricida</i> (Tc, 1), <i>Lygus lineolaris</i> (Ll, 1), <i>Nilaparvata lugens</i> (Nl, 9) and <i>Laodelphax striatellus</i> (Ls, 9). Branches of the genes from the same subclass are indicated by the same color.</p

Interaction between TYLCV C2 and AtRPS27A affects whitefly performance.

(A) In vitro GST pull-down assays. MBP or MBP- C2 fusion proteins were pull-down by GST or GST-AtRPS27A fusion protein. GST beads were washed and proteins were analyzed by SDS-PAGE western blot. Associated proteins were detected by anti-MBP antibody and gels were stained with Coomassie Brilliant Blue to monitor GST and GST fusion proteins. (B) In vivo BiFC analysis of C2 interaction with NtRPS27A. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein. Bars = 20 mm. (C) Subcellular localization of C2 and AtRPS27A. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein. Bars = 20 mm. (D) Survival rate of adult whitefly on wild type Arabidopsis plants and the transgenic C2 expressing Arabidopsis plants. Values are means± SE, n = 30. (E) Daily number of eggs laid by per female whitefly on wild type Arabidopsis plants and the transgenic Arabidopsis plants expressing C2. Values are means±SE, n = 30. Asterisks indicate significant differences between different treatments (P < 0.05; Student’s t test for all experiments). All experiments were repeated three times with similar results.</p

PaLCuCNV C2 interacts with RPS27A and promotes the performance of whitefly.

(A) In vitro GST pull-down assays. MBP or MBP-PaL-C2 fusion proteins were pull-down by GST or GST-NtRPS27A fusion protein. GST beads were washed and proteins were analyzed by SDS-PAGE western blot. Associated proteins were detected by anti-MBP antibody and gels were stained with Coomassie Brilliant Blue to monitor GST and GST fusion proteins. (B) In vivo BiFC analysis of PaL-C2 interaction with NtRPS27A or ubiquitin. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein which is located in nucleus. Bars = 20 mm. (C) Subcellular localization of PaL-C2, NtRPS27A and ubiquitin. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein. Bars = 20 mm. (D) Survival rate of adult whitefly on control and PaLCuCNV-infected tobacco plants. Values are means±SE, n = 30. (E) Daily number of eggs laid by per female whitefly on control empty-vector-inoculated and PaLCuCNV -infected tobacco plants. Values are means±SE, n = 30. (F) Survival rate of non-viruliferous and viruliferous adult whiteflies on cotton plants. Values are means±SE, n = 30. (G) Daily number of eggs laid by per non-viruliferous and viruliferous adult female whiteflies on cotton plants. Values are means±SE, n = 30. Asterisks indicate significant differences between different treatments (P < 0.05; Student’s t test for all experiments). All experiments were repeated three times with similar results.</p

Localization of C2 and NtRPS27A and their interaction.

(A) Structure of RPS27A. (B) Interaction between SH2-C2 and NtRPS27A in the yeast two-hybrid system. Yeast strain Y2H Gold co-transformed with the indicated plasmids was spotted on synthetic medium SD-Leu-Trp-His with x-α-gal and 2 mM 3-amino-1,2,4-triazole. The empty vectors pGBKT7 and pGADT7 were used as negative controls. (C) In vivo BiFC analysis of SH2-C2 interaction with NtRPS27A. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein that is located in Nuclei. Bars = 20 mm. (D) and (E) In vitro GST pull-down assays. MBP or MBP-SH2-C2 fusion proteins were pull-down by GST, GST-NtRPS27A, GST-ubiquitin or GST-ubiquitin32-76 fusion proteins. GST beads were washed and proteins were analyzed by SDS-PAGE western blot. Associated proteins were detected by anti-MBP antibody and gels were stained with Coomassie Brilliant Blue to monitor GST and GST fusion proteins. (F) Subcellular localization of SH2-C2 and different segments of NtRPS27A. Nuclei of tobacco leaf epidermal cells were marked with a RFP fusion protein that is located in Nuclei. Bars = 20 mm.</p