Fig. 4. Inability of recombinant TGFB2 and GDF9 to affect Pfkp and Ldha mRNA levels in cumulus cells. Although recombinant TGFB2 and GDF9 promoted cumulus expansion and recombinant GDF9 suppressed Lhcgr mRNA expression in mural granulosa cells, they did not promote expression of Pfkp and Ldha mRNA in OOX cumulus cells. OOX cumulus cells were treated with recombinant TGFB2 (A,B), GDF9 (D,E) or oocytes (2 oocytes/µl) and (A,D) degree of cumulus expansion and (B,E) expression levels of Pfkp and Ldha mRNA in OOX cumulus cells were examined. (C) Mural granulosa cells were cultured with recombinant GDF9 or oocytes (2 oocytes/µl), and Lhcgr mRNA expression in the mural granulosa cells was examined. Mean±s.e.m. The values indicated by different letters (a, b and c) are significantly different (P<0.05).