Non-human Primate Schlafen11 Inhibits Production of Both Host and Viral Proteins
Fig 2
Primate versions of Schlafen11 differentially inhibit retroviral protein production.
Plasmids encoding Schlafen11-V5 from the indicated primate species, or a V5-tagged chloramphenichol acetyltransferase (Chlor) gene as a negative control, were co-transfected into 293T cells along with plasmids encoding (A) a nearly full-length HIV clone (pNL4-3.Luc.R+E-), (B) HIV-1 Gag-Pol-RRE and HIV-1 Rev, (C) MLV Gag-Pol, (D) FIV Gag-Pol. Immunoblotting was used to monitor protein production of chloramphenichol acetyltransferase (Chlor-V5), Schlafen11-V5, GAPDH, and viral proteins. Panels A-D are representative blots from 3 or more experimental replicates. (E) Bands from panels A-D were quantified to show the relative effect of each Schlafen11 homolog. Each experiment was normalized to human Schlafen11. As increasing Schlafen11 activity as seen, the color of the box changes linearly along the blue to red color spectrum (see materials and methods for quantification procedure). (F) Viruses were packaged in 293T cells in the presence or absence of Schlafen11. Increasing amounts of the plasmids necessary for virus packaging were co-transfected along with a constant amount of plasmid expressing human, gibbon, or marmoset Schlafen11 (or Chlor as a negative control). The resulting virions were then used to infect 2.5x105 293T cells and the percentage of infected cells was scored by flow cytometry (GFP+). These data are representative of two independent experiments. Herein, “Chlor” is used as an abbreviation instead of the standard “CAT,” since the latter is also the name of another mammal and could therefore cause confusion.
