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Fig. 1. Oocyte stimulation of granulosa cell DNA synthesis - comparison with GDF9 and BMP6. (A) Mural GCs were cultured with increasing numbers of oocytes per well or increasing doses of recombinant BMP6 or GDF9 and assessed for [³H]thymidine incorporation after 24 hours. Points are means ± s.e.m. from triplicate wells and are representative of three replicate experiments. (B) Mural GCs were cultured alone or together with DOs (125 DOs/125-µl well) for 6 hours, RNA was extracted, reverse transcribed and subjected to real-time RT-PCR for Ccnd2. ^*mRNA levels significantly increased with oocyte co-culture. (C) Mural GCs were cultured alone, treated with BMP6 (50 ng/ml) or co-cultured with 16 oocytes per well, each with and without a neutralising dose (20 µg/ml) of a BMP6 monoclonal neutralising antibody (NAb) or a control IgG (20 µg/ml). Columns represent means ± s.e.m. from triplicate wells, representative of three replicate experiments. BMP6 NAb had no significant effect (P>0.05) on oocyte-stimulated GC DNA synthesis.

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