Figure 3
From: Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins
Indirect immunofluorescence assays of CCHFV glycoproteins after transfection of different expression plasmids. BHK-21 and 293T cells were transfected with CCHFV glycoprotein expression plasmids. Twenty-four hours post transfection cells were fixed and stained with CCHFV-specific or anti-HA antibodies. A: Cells fixed with methanol/acetone allow analyses of intracellular proteins. Polyclonal antibodies against CCHFV GC were used for GC detection (a, f). CCHFV GN expression from two different CMV-driven expression plasmids was analyzed using anti-HA tag antibodies (b, c, g, h). GN and GC expressed from the GPC were studied using polyclonal anti-CCHFV antibodies (d, i) as well as specific antipeptide antibodies against GN and GC (data not shown). CCHFV-infected cells served as controls (e, j). B: Cells fixed with paraformaldehyde were analyzed for CCHFV G expression on cellular surfaces. GC was stained using anti-CCHFV GC antibodies, GN with anti-HA tag, and mature CCHFV proteins derived from the GPC with anti-CCHFV antibodies. No clearly visible staining correlates with no detectable surface expression.