Fig. 2.

Tumor cell (TC) transmigration locations, transmigration rates and corresponding permeability of endothelial cell (EC) monolayers to BSA. Confocal images showing TC (red) adhesion to EC junctions (ZO-1, green) (A), TC transmigrating at the EC body (nucleus, blue) (B), at the joint between two ECs (bi-joint) (C), at the joint between three ECs (tri-joint) (D) and TC transmigrated to the basal side of an EC monolayer (E). Summary of locations of total transmigrating TCs across bEnd3 (mouse brain microvessel endothelial cell) and RLMEC (rat lung microvessel endothelial cell) monolayers out of 9 samples (3 samples each at 4, 6, and 8h) for each EC (F). * p < 0.05. Values are means ± SE. Numbers of adherent, transmigrating and transmigrated TCs at 4, 6, and 8h on/across bEnd3 (G) and RLMEC (H) monolayers. * p < 0.05. Values are means ± SE. Comparison of the number of effective transmigrated TCs across bEnd3 and RLMEC monolayers at 4, 6 and 8 h (I). The number of effective transmigrated TCs = the number of transmigrated TCs + 0.5 × the number of transmigrating TCs. * p < 0.05, comparison between different EC monolayers at the same time. ^$ p < 0.05, comparison between 6 h or 8 h with 4 h; ^@ p < 0.05, comparison between 8 h and 6 h, for each EC monolayer. Values are means ± SE. Comparison of the number of effective transmigrated TCs (green filled circles) and the permeability to BSA (red filled circles) at 4, 6 and 8 h for bEnd3 (J) and RLMEC (K) monolayers. The permeability of the control group without adding TCs is plotted with the blue filled circles for each EC monolayer. * p < 0.05, compared with the control permeability at the same time; ^$ p < 0.05, comparison between 6 h or 8 h with 4 h; ^@ p < 0.05, comparison between 8h and 6h, for each EC monolayer. Values are means ± SE. For all the measurements, n = 3 samples at each time for each EC.