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. 2021 May 4;9(5):457. doi: 10.3390/vaccines9050457

Figure 3.

Figure 3

Ability of R9-HuscFvs in inhibition of VLP cellular entry (neutralizing activity). (A) Ebola-VLP-based system used in this study. VLP consists of EBOV GP on the surface and VP40 matrix proteins fused with beta-lactamase (bla), which was produced by co-expression of EBOV GP and VP40-bla. Entry of VLP into cells were monitored by measuring bla activity, which cleaved cell-permeable fluorescence resonance energy transfer (FRET) substrate CCF2-AM (green color, emission wavelength at 530 nm), yielding the product (blue color, emission wavelength at 460 nm). (B–E) HEK293T cells were spinoculated with VLP, pre-incubated with indicated R9-HuscFvs (30 μg/mL final concentration in wells), the equivalent volume of diluent (diluent), or boiled VLP (mock infection). Percent neutralization (inhibition of cellular entry) was calculated as described in Materials and Methods and is shown in (B,D). Representative images of experiments in (B,D) are shown in (C,E), respectively. Results in (B,D) are shown as mean ± standard error of three independent experiments in triplicates. *, p < 0.05; **, p < 0.01; NS, not significantly different.