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Fig. 4. Effects of conditioned media derived from COS7 cells expressing hTeM, shTeM or shChM-I on HUVEC tube formation. (A) HUVECs were suspended in a mixture of culture media from COS7 cells transfected with the indicated cDNAs and fresh EBM medium at a ratio of 1:1. The cells were then plated on growth factor-reduced Matrigel in the presence of VEGF (20 ng/ml). Representative phase-contrast microscopic images after 6 hours incubation are shown. HUVECs suspended in fresh medium derived from wild-type untransfected COS7 cells and in medium collected from either mock- or hTeM-transfected cultures organized themselves into tubular networks. By contrast, HUVECs suspended in media derived from either shTeM- or shChM-I-transfected COS7 cells did not form tubular structures. Scale bar: 100 µm. (B) Total tube length per field was measured using image processing and analysis software. The data represent the mean±s.d. from three independent experiments. *P<0.01 versus HUVECs suspended in the EBM medium.
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