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. 2018 Mar 12;9:1029. doi: 10.1038/s41467-018-03432-4

Fig. 7.

Fig. 7

Infectivity of VSVΔG-GFP virus pseudotyped with mutated G in various cell types. a Generation of VSVΔG-GFP virus pseudotyped with VSV G mutants. Transfected HEK-293T expressing mutant G were infected with VSVΔG-GFP pseudotyped with VSV G WT. After 16 h, VSVΔG-GFP virions pseudotyped with mutant VSV G were harvested. b, c Incorporation of WT and mutant G in VSV∆G-GFP viral particles. VSVΔG-GFP pseudotyped with the WT VSV G was used to infect HEK-293T (MOI=1) transfected with indicated mutants. After 16 h, viral supernatants were analyzed by Western blot (using an anti-VSV G and an anti-VSV M antibody) (b). For each mutant, the G/M ratio was compared to that of WT normalized to 1. The mean of 3 independent experiments is shown with error bars representing the SD (c). d Infectivity of VSV∆G-GFP pseudotyped with WT and mutant glycoproteins. VSVΔG-GFP pseudotyped with WT VSV G was used to infect HEK-293T previously transfected with the indicated mutated glycoprotein (MOI 1). VSVΔG-GFP viruses pseudotyped by WT or mutant glycoproteins were used to infect indicated cell lines during 4 h; the percentage of infected cells was determined by counting GFP expressing cells by flow cytometry. Data depict the mean with SD for three independent experiments. Above each bar, the reduction factor of the titer (compared to VSVΔG-GFP pseudotyped by WT G which was normalized to 1) is indicated. e Sequence alignment of human (h) and drosophila (d) CR domains having characteristics similar to human LDL-R CR2 and CR3. Besides scaffolding residues of the CR fold (i.e., the 6 cysteines, labeled by stars, and the four acidic residues—I, II, III, IV—forming the calcium octahedral cage), we imposed (i) the absence of insertion or deletion in the contact region, (ii) an aromatic residue corresponding to W66 of CR2, and (iii) an amidic or an acidic amino acid in place corresponding to residue Q71 of CR2. PGBM: Basement membrane-specific heparan sulfate proteoglycan core protein. Vit.Rec: Putative vitellogenin receptor. Numbering of residues corresponds to that of the mature protein for CR2 and CR3 and to the precursor in all other instances (i.e., including the signal peptide)