Fig. 4

PAI-1 derived from CAFs is required to induce EndoMT in LECs. A Expression profiles of cytokines in NF-CM and CAF-CM. B Overlap between lymphangiogenesis-related and EMT-related proteins. C The expression of the four significant cytokines was analysed by qRT-PCR. D The secretion of PAI-1 from primary fibroblasts was analysed by ELISA. E HDLECs were treated with CM in the presence of control anti-IgG (represented as “-”; GeneTex, GTX35009, 20 μg/ml) or anti-PAI-1 antibody (represented as “ + ”; GeneTex, GTX79745, 20 μg/ml). Cell lysates were subjected to Western blotting with antibodies against the indicated proteins. F Representative micrographs of migration assays (upper panels), tube formation assays (intermediate panels) and transendothelial assays (lower panels) using HLECs pretreated with the indicated treatments. G The average number of migrated cells per field was calculated. H The average tube length per field was calculated. I The number of transmigrated SiHa-mCherry cells on the bottom side of the HDLEC monolayer was quantified. Data are presented as the mean ± SD of three independent experiments. *P < 0.05