Fig 5. Passive transfer experiment for B12L.

a) Clinical scores were recorded at 3 h, 6 h, 9 h, 24 h, 30 h, and 48 h after intravenous administration of B12L or saline (n = 3, each group, mean ± SD). Since the animals administered B12L at a dose of 3 mg/kg were moribund and euthanized 30 h after dosing, their clinical scores were recorded as ‘4’ at 48 h. b) Body weights were measured at 0 h, 24 h and 48 h after administration (n = 3, each group, mean±SD). We could not measure the body weights in the group administered 3 mg/kg of B12L at 48 h, because they were moribund and euthanized at 30 h after administration. c) Representative images of complement deposition in control rats (low magnification [A, B, and C], high magnification [G, H, and I]) and B12L-treated rats (low magnification [D, E, and F], high magnification [J, K, and L]) are shown. C3 (D and J, green) and α-Btx (E and K, red) signals were colocalized in neuromuscular junctions (NMJs) (F and L, merged) at 8 h after administration of B12L at a dose of 1.5 mg/kg. Scale bar = 50 μm. d) The amount of nAChRs in NMJs was calculated by imaging analysis. The graph shows the signal ratio of α-Btx (red) colocalized with SV2A (green) compared to SV2A signal alone in NMJs at 48 h after administration of saline or B12L at a dose of 1.5 mg/kg (n = 4, each group, mean±SD).