Fig. 8. Effect of VEGFR-3 inhibition on the transmigration of BAE cells in response to VEGF-C_NC. BAE cells were induced to transmigrate through 8 µm polycarbonate filters by the addition of 5 ng/ml ligand to the bottom well; transmigrating cells were stained with Hoechst stain, imaged under epifluorescence using a 20x lens and counted using automatic image analysis (see Materials and Methods). (A) Chemotaxis towards VEGF-C_NC is inhibited in a dose-dependent manner by mAb hF4-3C5 () whereas mAb IMC-1C11 () has no effect. Combination of the two antibodies showed no additive effect (). Results are shown as the means±s.e.m. (Inserts) Examples of transmigrated cells stained by Hoechst and used in automated cell counting; (A1) no antibody; (A2) 100 nM each of hF4-3C5 and IMC-1C11. (B) Saturating amounts of VEGFR-2-AP were mixed with various amounts of mAbs and added to 96-well plates coated with VEGF-C_NC. Bound receptor was measured using AP activity. Blocking of the receptor binding is evident with the mAb IMC-1C11 () but not with the irrelevant mAb IMC-C225 ().