Characterization of iLECs. (A-D) Immunofluorescent staining of iLECs (A,C) and human dermal microvascular endothelial cells (HDMECs; B,D) for PROX1 (green) and β-catenin (red; A-B) or podoplanin (green) and 4,6-diamidino-2-phenylindole (DAPI; blue; C-D). HDMECs are mixed dermal endothelial cell populations containing both LECs and BECs. BECs are PROX1-negative and express higher levels of β-catenin.9 Bars represent 50 μm. (E) Northern blotting of mRNA from 293T cells, and BECs, dLECs, and iLECs from 2 different persons hybridized with probes for the indicated transcripts. The 28S ribosomal RNA is shown as loading control. (F) Increased VEGFR-3 phosphorylation on stimulation of iLECs with VEGF-C. (G) Activation of ERK1/2 and Akt phosphorylation after stimulation of iLECs with VEGF-C or VEGF-C156S. Total ERK1/2 and Akt levels are shown as control. (H) VEGF-C and VEGF-C156S induce iLEC migration. The results are shown as mean ± SEM. (I) VEGF-C and fibronectin (FN) enhance survival and proliferation of iLECs and dLECs. The results are shown as mean, relative to day 0, ± SEM.
