Transient, local blood vessel permeability events accompany intravasation at the TMEM. A, IVM time lapse of 155-kDa dextran–TMR extravasation and tumor.

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TMEM-mediated vascular permeability is transient and localized.

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Transient, local blood vessel permeability events accompany intravasation at the TMEM. A, IVM time lapse of 155-kDa dextran–TMR extravasation and tumor cell intravasation. Transient, local blood vessel permeability events accompany intravasation at the TMEM. A, IVM time lapse of 155-kDa dextran–TMR extravasation and tumor cell intravasation. TMEM, white box. Blood vessel permeability sites (white arrowheads) and intravasating tumor cell (TC; yellow dashed line, 9′). Clearance of dextran and decrease of CTCs at 30′. Scale bar, 50 μm. At 9′ and 30′, TMEM tumor cells and macrophages are added in false color to increase visibility after bleaching. B, isolated 155-kDa dextran–TMR channel from A. Red arrowheads mark dextran extravasation (white). Dashed red line, luminal side of the endothelium. C, isolated tumor cell channel from A. Yellow arrow marks site of intravasating tumor cell (yellow dashed line) at the TMEM. White dashed line marks the luminal surface of the endothelium. Red box, region adjacent to the TMEM with elevated CTCs. D, single time point of tumor cell intravasation (yellow dashed line) by time-lapse IVM. Scale bar, 50 μm. E, 3D reconstruction of time-lapse IVM from D of tumor cell intravasation at the TMEM. Transmigrating tumor cells (individually numbered, dashed white lines) are isolated from other cell types for clarity with time in minutes from start (J0′) to end of transmigration (J3′). The luminal endothelial surface is outlined in a pink dashed line. Extravascular dextran (red) at the TMEM indicated with a yellow arrowhead and outlined in a yellow dashed line. F, frequency of blood vessel permeability events in the presence of the TMEM or away from the TMEM in 100-μm windows (see Supplementary Fig. S4; n = 16; **, P = 0.0034). G, frequency of tumor cell intravasation events in the presence of the TMEM or away from the TMEM in 100-μm windows (n = 16; **, P = 0.0012). H, quantification of extravascular dextran intensity and CTC area at the TMEM over time from A. , extravascular dextran; , CTC. Allison S. Harney et al. Cancer Discovery 2015;5:932-943 ©2015 by American Association for Cancer Research