Figure 9.

Molecular characterization of h-TERT-HDMVEC. A: hTERT-HDLEC integrin and MMP expression were assessed by RT-PCR. The total cellular RNA, isolated from confluent hTERT-HDLEC monolayers (at passage 26) cultured in EGM-2MV Bullet kit medium, was reverse-transcribed and amplified by PCR. The specific conditions and primers pairs are indicated in Table 1. Equal amounts of PCR products were analyzed on a 2% agarose gel. Marker (123-bp DNA ladder), hTERT-HDLEC + RT (lane 1), hTERT-HDLEC − RT (lane 2), human liver + RT (lane 3), H₂O (lane 4). The expression of αvβ3 and αvβ5 integrin complexes was confirmed by FACS analysis (data not shown). B and C: Zymographic analysis of HMEC-1 and hTERT-HDLEC. Cells were cultured in serum-free media lacking FGF-2 or VEGF-A. Confluent monolayers in 35-mm tissue culture dishes were left untreated (control) or exposed to 100 ng/ml VEGF-A or 100 ng/ml VEGF-C for 15 hours. Supernatants were analyzed by gelatin (B) or casein (C) zymography. uPA activity was confirmed by its inhibition with amiloride (data not shown).