Rhesus macaque IFITM3 exerts antiviral activity.

<p>293T cells expressing human (hu) and rhesus macaque (rh) IFITM3 or chloramphenicol-acetyltransferase (CAT) as negative control were transduced with MLV particles encoding luciferase and harboring the indicated viral glycoproteins. Luciferase activities in cell lysates were determined at 72 h post transduction. The average of three to six independent experiments is shown. Luciferase activities measured in control cells were set as 1. Error bars indicate standard error of the mean.</p

Overview of sequence variations identified in the <i>Macaca mulatta IFITM3</i> locus.

<p>The <i>Macaca mulatta IFITM3</i> locus and positions of primers are shown as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172847#pone.0172847.g001" target="_blank">Fig 1</a>. The positions of polymorphisms are marked by stars and the exact nucleotide position (relative to the start codon) and the detected variations are given.</p

Polymorphism-specific rank-based tests (p values) of genotype differences in virus load.

<p>Polymorphism-specific rank-based tests (p values) of genotype differences in virus load.</p

Frequency of polymorphisms.

<p>Frequency of polymorphisms.</p

Polymorphism-specific log-rank tests (p values) of genotype differences in AIDS-free survival.

<p>Polymorphism-specific log-rank tests (p values) of genotype differences in AIDS-free survival.</p

Description of the cohorts.

<p>Description of the cohorts.</p

<i>IFITM3</i> alleles and viral load at set point in SIV-infected rhesus macaques.

<p>For each polymorphism, the virus load at set point (20 weeks post infection) was compared between the different genotypes. The polymorphisms are indicated above each diagram along with the p-values obtained by Mann-Whitney or Kruskal-Wallis test. Virus load is shown as RNA copies/mL plasma.</p

Tunneling Rearrangement of 1‑Azulenylcarbene

1-Azulenylcarbene was synthesized by photolysis of 1-azulenyldiazomethane in argon or neon matrices at 3–10 K. The highly polar singlet carbene is only metastable and undergoes a tunneling rearrangement to 8-methylene-bicyclo[5.3.0]­deca-1,3,5,6,9-pentaene. After substitution of the 4 and 8 positions with deuterium, the rearrangement is completely inhibited. This indicates a very large kinetic isotope effect, as expected for a tunneling reaction

Immunofluorescence analysis of HFF cells infected with recombinant viruses.

<p>The co-localisation of viral protein and EYFP in HFF infected with the recombinant virus was detected by fluorescence microscopy at 40× magnification. HFF were infected with TB4-IE2-EYFP (<b>A</b>), TB4-UL83-EYFP (<b>B</b>) or TB4-UL32-EYFP (<b>C</b>) with MOI 1 and fixated at 6, 24, 48 and 72 hpi as indicated on the left side. Cells were stained with mouse monoclonal antibodies directed against IE1–2 (E13) (A), ppUL83 (B) or ppUL32 (XP-1) (C) and Alexa-Fluor555 conjugated secondary antibody. Single channel recordings of DNA (Dapi), EYFP and the viral protein portion (red staining with Alexa-Fluor555) are shown in the left three columns; merged images are in the rightmost column. The time after infection is indicated on the left side.</p

Testing measurement of antiviral agents.

<p>HFF infected in 96-well plates and fluorescence was measured in live cells by fluorescence spectrometry or detected by live microscopy at 10-fold magnification. (<b>A</b>) The relationship of relative fluorescence intensity (RFI) and multiplicity of infection is shown in the left panel for cells infected with the indicated viruses for 8 days. The panels on the right show cells infected with TB4-IE2-EYFP (5 dpi) and TB4-UL83-EYFP or TB4-UL32-EYFP (8 dpi) at the indicated MOI. (<b>B</b>) Measurement of neutralizing activity using TB4-IE2-EYFP. Virus was incubated with different dilutions of Flebogamma^R (5%) (upper row) or supernatant from hybridoma cell lines producing antibodies directed against anti-gH (14-4B) (middle row) or anti-gB (27–39) (bottom row). HFF were infected at MOI 0.1 and relative fluorescence intensities recorded at 9 dpi are shown in the panels on the left. Corresponding microscopic images are shown in the panels on the right.</p