Fig. 1.

CD reduces LCMV GP expression levels. A) Amino acid sequence of LCMV GP CD. CD LCMV GP amino acid residues are indicated in red. Methionine (M) and tryptophan (T) residues in LCMV GP, as well as amino acids already associated with deoptimized codons are indicated in black. Underlined (solid) amino acids represent the stable signal peptide (SSP). No underlined region represents GP1. Underlined (dotted) amino acids represent GP2. Box represents the GP transmembrane domain. Amino acids highlighted in gray represent the GP cytoplasmic tail. B) Mutations in LCMV GP CD. Number of nucleotide mutations and the percentage (%) of CD amino acids are indicated. C-E) LCMV GP CD expression levels in transfected cells. Human 293 T cells were transiently transfected with pCAGGS expression plasmids encoding LCMV GP WT or GP CD and at 48 h p.t. GP expression was assessed by IFA (C), FACS (D) and WB (E) using the LCMV GP mouse monoclonal antibody 83.6 (GP2). Empty (E) plasmid was included as negative control (C–D). IFA, FACS and WB results correspond to representative of three independent transfection experiments. (C) Scale bar=100 μm. (D) Blue line: mock-transfected cells. Black line: cells transfected with LCMV GP WT. Red line: cells transfected with LCMV GP CD. (E) GAPDH expression levels were used as loading controls.