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Thermolysin treatment results in greater extents of fusion between cells.

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posted on 2016-01-18, 14:25 authored by Ruben M. Markosyan, Chunhui Miao, Yi-Min Zheng, Gregory B. Melikyan, Shan-Lu Liu, Fredric S. Cohen

(A) schematic of the experimental protocol is shown above the bar graph. E, effector cells; T, target cells, Th, thermolysin. Bar graph: Fusion of thermolysin-treated effector cells expressing EBOV GP (columns 1 and 2, dark red) was greater than for untreated cells (columns 3 and 4, dark yellow). For both thermolysin-treated and non-treated cells, a 10-min pH 5.7 pulse applied at room temperature augmented fusion, measured after an additional 2 h incubation at neutral pH. For each condition, at least 7 experiments were performed. Typical images used to obtain the data of the bar graph are shown on the right: in top images, cells were treated with 200 μg/ml thermolysin; in bottom images, cells were not treated. Cells that have fused are marked by arrows. (B) The kinetics of fusion for thermolysin-treated (dark red squares) and untreated (dark yellow circles) effector cells. Cleaving EBOV GP by thermolysin speeds fusion kinetics, but extents of fusion are the same for treated and untreated cells after a pH 5.7 pulse at 10°C is followed by a 4 h reneutralization. * p <0.05; *** p < 0.001.