Mab3552

C57BL/6 mice were subcutaneously injected with CMT93 cells with or without stable Dragon over-expression, followed by intraperitoneal injection of anti-BMP2/BMP4 antibody (MAB3552, R&D, Minneapolis, MN, USA) or IgG₁ isotype control (MAB002, R&D, Minneapolis, MN, USA) at a dose of 2 μg/g body weight, once every 2 days, for up to 22 days.

Cell signalling inhibitors were applied directly to cortical cultures (1 h before exposure to CM) or indirectly via the in vitro model, the BeWo barriers (at exposure to altered oxygen) to see the effect on CM. Map kinase inhibitor SB203580 (20 μM; InviviGen), epidermal growth factor receptor (EGFR) inhibitor PD168393 (20 μM; Santa Cruz Biotechnology), Noggin (200 ng/ml; StraTech), Gremlin (Gremlin-2, 500 ng/ml; Sigma) and Chordin (chordin-like 1, 500 ng/ml; Sigma) were tested. Inhibitors were chosen to block signals in the Smad (Noggin, Chordin, and Gremlin), mTOr-Akt-PI3k (PD168393) and 38-MAPK (SB203580) pathways in an attempt to isolate which if any signalling pathway was being affected. Where appropriate Bone Morphogenetic Protein (BMP) binding antibody (R&D Mab3552) was applied to cultures at a dose of 10 µg/ml 1 h before exposure to CM.