Figure 2.

SVMP and SVSP cleave human complement proteins. Human C3, C4 and C5 purified proteins (2.5 µg) were incubated with sterile saline or N. annulifera venom (NaV - 2.5 µg) with or without metallo- (10 mM) and serine-proteinases (10 mM) inhibitors for 30 minutes at 37°C. After this period, reactions were stopped and C3 (A), C4 (B) and C5 (C) cleavage evaluated by SDS-PAGE and silver stanning. Images presenting in panels a-c were grouped and/or spliced (the original gels are presented as supplementary material – Figure S5 ). C3a (D), C4a (E) and C5a (F) anaphylatoxins generation in these reactions were determined by CBA. Panels (A–F) represents five different experiments. Data are means ± SEM of three independent experiments ***p ≤ 0.001 (two-tailed t-test). ^### indicates a significant difference between complement purified proteins treated with N. annulifera venom + Vehicle and N. annulifera venom + Protease inhibitors. ^##p ≤ 0.01, ^###p ≤ 0.001. ns = non-significant.