Summary of the Structural Organization and Different Conformations of the Flavivirus Envelope Protein E (obtained the kind permission from the copyright holder to reproduce figures that have previously been published on 51)

<p><b>Copyright information:</b></p><p>Taken from "Does Japanese encephalitis virus share the same cellular receptor with other mosquito-borne flaviviruses on the C6/36 mosquito cells?"</p><p>http://www.virologyj.com/content/4/1/83</p><p>Virology Journal 2007;4():83-83.</p><p>Published online 6 Sep 2007</p><p>PMCID:PMC2075493.</p><p></p> (A) Schematic top view of the organization of the sE protein dimer as present at the surface of mature virions, color-coded according to the three domains (DI, DII, and DIII). The fusion peptide (FP) is indicated in orange. (B) Crystal structure (top view) of the TBEV E ectodomain (termed "sE") dimer. (C) Schematic side view of the DV E dimer at the surface of mature virions, with the "stem" and TM C-terminal polypeptide segments (missing in the truncated sE form) indicated in green. The viral lipid bilayer is illustrated with lipids belonging to the outer and inner leaflets colored blue and pink, respectively. Cryo-electron microscopy 3D reconstructions have shown that the stem forms two α-helices (H1 and H2) lying on the viral membrane, followed by the two transmembrane (TM) segments. (D) Schematic representation illustrating the proposed organization of full-length DV E in its postfusion conformation. In this model, the α-helices of the stem interact with the body of the trimer, in the grooves between adjacent, parallel DIIs. The lipid bilayer as well as the stem and TM segments is drawn as in (C)

Supplemental Material, Supplemental_Figure_legends - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Supplemental_Figure_legends for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

Supplemental Material, Suppl_Figure_3 - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Suppl_Figure_3 for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

Different sets of transcription factors bind to <i>Prnd</i> promoter in N2a and GC-1 spg cells determined by gel-shift analyses with the nuclear extract (NE) from GC-1 spg and N2a cells.

<p>A. USF binds to E-box in both N2a and GC-1 cells; B. Brn-3 binds to the G and T rich regions −1295/−1278 and −1215/−1199 in GC-1 spg but not in N2a cells; C. Sp1 binds to GC-box in GC-1 cells and Sp3 binds to GC-box in N2a cells. Combination of the oligonucleotides and nuclear extracts used in assays are indicated by colorful circles above the radiogram.</p

Supplemental Material, Ding_Supp_Fig_2_final - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Ding_Supp_Fig_2_final for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

Dpl induces ROS accumulation and acute cell death in N2a but not in GC-1 spg cells.

<p>A. Dpl induces ROS accumulation. N2a (a) or GC-1 spg cells (b) in 96-well plates were incubated with 50 µM DCFH-DA for 45 min. After wash, cells were then incubated without or with MgCl₂ (100 µM), CuCl₂ (100 µM) or the purified mouse Dpl protein (20 µg/ml) for 1, 2 or 4 h. After wash, DCF fluorescence was determined at an excitation of 485 nm and emission of 538 nm by a microplate-reader. The readings of DCF fluorescence of each test group were normalized against that of the DCFH-DA control group and expressed as relative-fold change. Bars are means ± SD of 3 independent experiments. Student <i>t</i> test was used for statistic analyses (*, <i>p</i><0.05; **, <i>p</i><0.01). B. Dpl induces acute cell death. N2a (a) or GC-1 spg cells (b) in 96-well plates were incubated without or with MgCl₂ (100 µM), CuCl₂ (100 µM), or the purified Dpl (20 µg/ml) for 0, 1, 2, or 4 h. The cell growth curves were determined by MTT assay. Bars are means ± SD of 3 independent experiments. Student <i>t</i> test was used for statistic analyses.</p

Dpl induces ATM-dependent bindings of Sp1 and p53 to the <i>Prnp</i> promoter.

<p>A. Schematic locations of the putative binding sites of Sp1 and p53 in the mouse <i>Prnp</i> promoter. B. ATM-dependent bindings of Sp1 and p53 to the <i>Prnp</i> promoter. N2a cells were mock- (lanes 2–5) or transfected with the control siRNA (lanes 6, 7) or siRNA to ATM (lanes 8, 9) for 48 h and then incubated with Dpl (20 µg/ml) for 2 h. Gel-shift analyses were then performed with nuclear extracts from cells. Combination of the oligonucleotides and the nuclear extract used in the assay are indicated by black circles above the radiogram.</p

The differential activates of <i>Prnd</i> promoter are measured in N2a and GC-1 spg cells.

<p>A. Schematic structure of the <i>Prnd</i> and its promoter construct: 5′-flanking region and exon-intron organization of the <i>Prnd</i> region. The numbers of restriction sites represent the distances from the mRNA start site. B. The constructs of the <i>Prnd</i> promoter in pTal-Luc reporter vector. The plasmid containing the <i>Prnd</i> promoter regions of −1863/+27, −940/+27 or −185/+27 was digested with BamH I+Hind III (a), Nhe I+Hind III (b) and Bgl II+Hind III (c), respectively, and then run on the 1% agarose gel. <b>C</b>. Transcriptional activities of the <i>Prnd</i> promoter. N2a and GC-1 spg cells were transfected with the different <i>Prnd</i> promoter constructs (B) in pTal-Luc vector. Promoter activities are expressed in % relative to the activity of the full <i>Prnd</i> promoter construct (−1863/+27) in GC-1 spg cells, which was set to 100%. Bars are means ± SD of 3 independent experiments. Student <i>t</i> test was used for statistic analyses.</p

Dpl induces differently molecular effects in N2a and GC-1 cells.

<p>A. Dpl induces apoptosis in N2a cells but not in GC-1 spg cells. GC-1 spg (lanes 1-3) or N2a cells (lanes 4-6) were mock-transfected (lanes 1, 4) or transfected with pcDNA3 (lanes 2, 5) or pcDNA3-Dpl (lanes 3, 6) for 72 h. Cell lysates were subjected to Western blots with anti-Dpl mAb 1A9 (a) or anti-caspase-3 antibody (b). B. Dpl induces the phosphorylation of p53 and p21 in N2a but not in GC-1 spg cells. GC-1 spg (lanes 1–4) or N2a cells (lanes 5–8) were incubated with the purified Dpl protein (20 µg/ml) for 0, 1, 2 or 4 h. Cell lysates were subjected to Western blots with anti-phosphorylated p53 at S15 antibody (a), anti-p21 antibody (b) and anti-β-actin antibody (c). The protein signals of phosphorylated p53 at S15 (p53-S15-P) (a, d), p21 (b, e) and phosphorylated p21 (p21-P) (b, f) were scanned, normalized to β-actin levels (c), and expressed as relative -fold change over signals in untreated GC-1 spg cells (a1) or N2a cells (b5, c5). Bars represent the mean ± S.D. of three independent experiments. ANOVA were used for statistic analyses (**, <i>p</i><0.01). C. Dpl induces the elevation of the full-lengh PrP^C in N2a cells but has no effect on the N-terminal truncated PrP^C in GC-1 spg cells. N2a (lanes 5–8) or GC-1 cells (lanes 1–4) were incubated without (lanes 1, 3, 5, 7) or with Dpl (20 µg/ml) (lanes 2, 4, 6, 8) for 1 h (lanes 1, 2, 5, 6) or 2 h (lanes 3, 4, 7, 8). Cell lysates were subjected to Western blots with anti-PrP mAb SAF-32 (a), SAF-70 (b), or anti-β-actin antibody (c).</p

Dpl and PrP are differentially expressed in N2a and GC-1 spermatogenic cells.

<p>A. The target epitope of monoclonal antibody 1A9 to Doppel (a) and SAF-32 or SAF-70 to PrP (b). B. Expressions of Dpl and PrP in N2a and GC-1 spg cells. Cell lysates of mouse N2a (lanes 1) or GC-1 spg cells (lanes 2) were subjected to Western blots detecting by anti-PrP mAb SAF-32 (a), SAF-70 (b), or anti-Dpl mAb 1A9 (c).</p