Figure 5
From: Neutralization of Junín virus by single domain antibodies targeted against the nucleoprotein
VHHs 1.2, 1.69, 2.96 and 2.3 recognize N derived from Candid# and XJCl3 strains. (a) Vero cells at 60% confluence were transfected with 200 ng of expression plasmids coding for N from Candid# (left) or XJCl3 (right) strains. At 48 h post-transfection, the cells were fixed, permeabilized and further incubated with 10 µM of VHHs followed by incubation with N-specific mAbs. Afterwards, cells were incubated with an anti-His antibody and finally incubated with the respective secondary antibodies. Immuno-staining without VHH (−VHH), without N-specific mAb (−N), with a VHH against Flu (VHH Flu) and with GPC-specific mAbs (GPC) were included as controls. Pictures are representative of 3 independent experiments. (b) Colocalization of the fluorescent signal derived from immuno-staining with the VHHs was determined in 30 cells that showed a positive signal with N-specific mAbs. The Pearson’s correlation coefficient (P) was calculated using the Coste’s algorithm provided in the Volocity software package. The scale bar represents 25 µm.
