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. 2016 Nov 17;11(11):e0166013. doi: 10.1371/journal.pone.0166013

Fig 1. Gn and Gc proteins are efficiently expressed from separate plasmids.

Fig 1

(A) 293T cells were transfected with empty pCAGGS plasmid or pCAGGS encoding the indicated viral glycoproteins with a C-terminal V5 antigenic tag. Glycoprotein expression was analyzed by Western blotting using a V5-specific monoclonal antibody (top panel). Detection of β-actin served as loading control (bottom). Unprocessed glycoprotein precursor proteins are marked with filled triangles while asterisks indicate processed Gc (SFTSV, RVFV, LACV) or GP2 (EBOV, LASV). A single representative blot is shown from which irrelevant lanes were excised. Similar results were obtained in three independent experiments. (B) The experiment was conducted as described for panel A but plasmids encoding both the Gn/Gc precursor and single proteins (all containing a V5 tag) were transfected. A single representative blot is shown from which irrelevant lanes were excised. Similar results were obtained in three independent experiments. (C) The experiment was conducted as described for panel B but cell lysates were treated with PNGaseF to remove N-glycans or Mock treated. As a control for successful removal of N-glycans, cells expressing the spike protein of MERS-CoV were PNGase F treated. The results were confirmed in three independent experiments.