Fig. 2

Expression of syEBOVgD106. a Diagram (not on scale) of pINT2CMV-syEBOVgD106 targeting vector delivering the tagged syEBOVgD106 ORF (syEBOV, orange; gD106, red), under the control of the CMV promoter (CMV, blue) and the bovine growth hormone polyadenylation signal (PA, yellow). CMV-syEBOVgD106 expression cassette is flanked by BoHV-4 TK homologous sequences (white). b Western immunoblotting of pINT2CMV-syEBOVgD106 transfected HEK 293T cells extracts. Lanes were loaded with different amounts of total protein cell extract (5, 10 and 20 μg); cells transfected with pEGFPC-1 served as negative controls (Mock). The peculiar immune-blotting banding pattern is the result of the syEBOVgD106 protein processing by furin and TACE proteases (c), as revealed by the anti gD106 tag monoclonal antibody directed against three predicted and detected peptides (1 uncleaved; 2 only cleaved by Furin protease; 3 cleaved by Furin and TACE proteases) (d)