Fig. 5. CD105 mRNA and protein expression in ODN-treated and untreated cells. HDMECs were cultured for 72 hours in normoxia in the presence or absence of ODN plus DMRIE-C, followed by exposure to hypoxia for 24 hours. CD105 protein levels were assessed by immunoblotting and FACS, and mRNA levels were assessed by northern blotting. (A) Immunoblotting revealed that CD105 protein was reduced by 58% in AS-ODN-treated cells (lane 2) compared with SC-ODN-treated (lane 3) and untreated cells (lane 1). The bar chart indicates the ratio of CD105 relative to CD31 signal intensity collected from three experiments (means±s.e.m.). (B) FACS analysis of cell surface CD105 indicated a 60% reduction in AS-ODN-treated (blue profile) compared with SC-ODN-treated (green profile) or untreated (black profile) cells. The red profile represents a negative control, where mAb E9 was substituted with pre-immunised mouse serum. The plot is a representative of five similar experiments. (C) Northern blotting of CD105 and GAPDH mRNA showed that CD105 mRNA was markedly degraded by AS ODN (lane 2) but not by SC ODN (lane 3). Lane 1 is mRNA extracted from untreated cells, and it has no evident alteration compared with lane 3. The bar chart shows CD105 mRNA, relative to GAPDH mRNA signal intensity, pooled from three experiments (means±s.e.m.).