Cmac dye

Confluent monolayer of ID8 cells were pre-treated with 5 µM of Compound C or DMSO for 18 h and were labelled with CellTracker™ Blue 7-amino-4-chloromethylcoumarin (CMAC) Dye (Invitrogen) for 30 min. C57BL6 mice were injected intaperitoneally with 1 × 10⁶ CMAC-ID8 cells. After four hours, mice were euthanized, omenta were collected in 6 well cell plates and gently washed once with PBS (n = 4 mice per group). After which, specimens were preserved in 70% ethanol. Fluorescence imaging was immediately conducted using Olympus IX-70 at 460 nm (blue) and visualized in 5 fields/organ at 100× magnification and analyzed using Image J software.

Before the microdevice was assembled, all tubes and PDMS plates were pre-treated with 75% ethanol for 12 h, and the entire system was then dried in oven at 60°C. The dried devices were exposed to ultraviolet light for 30 min. The porous membrane was coated with collagen type I hydrogel at 37°C. The Caco-2 cells (1 × 10⁵ cells/cm²) stained by blue cell-tracker (CMAC Dye, Invitrogen) were seeded on the porous PDMS membrane and incubated at 37°C for 3–4 h, allowing the seeded intestinal epithelial cells to attach to the membrane surface. Then, HUVECs (1 × 10⁵ cells/cm²), which were stained by green cell-tracker (CMFDA Dye, Invitrogen) and wrapped by type I collagen gel, were seeded onto the basal of the membrane. After vascular endothelial cell attached to the membrane, the microdevice was assembled and ran as described above.