Fig. 4.
Effect of a fibritin trimerization motif on the TMmod10 Env. a Cell lysates and supernatants from 293T cells expressing the EKNA variants of sgp140(−) or TMmod10, or these Envs with a C-terminal fibritin trimerization domain (sgp140(−) FT and TMmod10 FT, respectively), were analyzed on gels and Western blotted. A polyclonal goat anti-gp120 antibody was used to detect the Envs on the Western blots shown in this figure. Addition of the fibritin domain to the C-terminus of TMmod10 did not prevent Env expression but significantly diminished release of the TMmod10 FT glycoprotein from the cells. b The indicated soluble Envs were subjected to digestion with PNGase F or Endo Hf and Western blotted. c Antibody precipitation of the EKNA variants of sgp140(−), sgp140(−) FT, TMmod10 and TMmod10 FT Envs secreted into the medium of expressing cells is shown. All EKNA Env constructs contain the E168K + N188A changes that restore the PG9/PG16 epitopes [96–99]. Data are representative of those obtained in duplicate or two independent experiments